Bulletin of Botanical Research

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Comparison on Leaf Anatomical Structures betweenthe Female and the Male Plants of Eippophae rhamnoides L.

LI Na, GUO Xue-Min, MENG Yang

Bulletin of Botanical Research 2017, 37 (1): 1-7. DOI: 10.7525/j.issn.1673-5102.2017.01.001

Abstract （978）

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We observed the differences in leaf anatomical structure between the male and the female using paraffin method to investigate leaf secondary sex characteristics of Hippophae rhamnoides L.. Results showed that:(1)Both the male and the female leaves consisted of epidermis, mesophyll and veins. The epidermis was composed of a layer of cells with developed trichomes, including similar bulliform cells in upper epidermis; the differentiation of palisade tissue and spongy tissue was obvious in mesophyll. (2)There were more similar bulliform cells in upper epidermis, more ergastic substances in some parenchymal cells of phloem and below the phloem, more trichomes in lower epidermis in the female plant than in the male one; the male plant were significantly higher than the female in leaf thickness, thickness of leaf upper epidermis, thickness of palisade tissue, thickness ratio palisade tissue/sponge tissue, and had more developed vascular bundle than the female in main vein. These results suggested that there were obvious differences in leaf anatomical structure between the female and the male plants of the species, and the differences were the performances of secondary sex characteristics, also robust adaptive strategy formed in the long-term evolution, which might be good for reproduction of the species.

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Development of Gametophyte and Young Sporophyte of the Fern Aleuritopteris argentea

CHEN Si-Qi, JIAN Jin-Jing, ZHANG Yan, DAI Xi-Ling

Bulletin of Botanical Research 2017, 37 (1): 8-14. DOI: 10.7525/j.issn.1673-5102.2017.01.002

Abstract （943）

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Aleuritopteris argentea is a small ornamental ferns. We observed the developmental features of gametophyte and sporophyte of A.argentea cultured in Knop's medium and humus medium using microscopical techniques, and studied the optimum pH value for the gametophyte development. The results show that:(1)the spores are yellow-brown, tetrahedral, and trilete with reticulate ornamentations. Polar view is triangle circle and the equatorial view is nearly semicircular. The germination is Vittaria-type and the gametophyte development is Ceratopteris-type. The mature gametophytes are cordate. The archegonia and antheridia are leptosporangiate-type. No hairs are formed during gametophyte development. The above features of the gametophytes in the fern A.argentea are stable. (2)the enhancement the pH value in 7.0-9.0 is beneficial for spore germination and gametophyte development. (3)the spore and gametophytes develop on humus medium produced seedlings after 7-8 weeks. The viability of sporophytes obtain in this medium are strong with developed roots and easy to transplant. Percentage of seedling emergency reaches to 90%. Our investigation is benefit for the artificial propagation and evolution of the resource plant A.argentea.

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Cytological Study on Microspore Abortion in OT Hybrid Lily Cultivars ‘Cocossa’

WU Jie, ZHAO Yu-Ping, HE Xiang-Feng, WANG Wen-He, LENG Ping-Sheng, ZHANG Ke-Zhong

Bulletin of Botanical Research 2017, 37 (1): 15-22. DOI: 10.7525/j.issn.1673-5102.2017.01.003

Abstract （846）

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We studied meiosis process and tepetum changes of OT hybrid lily cultivars ‘Cocossa’ using aceto-carmine dyeing method combined with grilled slices paraffin sections. When the bud grew to 2.5-4.5 cm, the pollen mother cell was in the period meiosis. There were some abnormalities appearances of meiosis:chromosome ring appeared in metaphase Ⅰ, chromosome bridges and lagging chromosomes ppeared in anaphaseⅠ, unequal division and micronuclei appeared in telophaseⅠand Ⅱ, it could also be observed 2 split-phase in the same plant, the same flower bud and even the same anther, that showed meiosis had not synchronized. During the telophaseⅡ and mononuclear pollen period, the tapetum delayed degradation, which led to multinuclear pollen, abnormal pollen and microspore infertility. The meiotic abnormalities and abnormal tapetum degradation could common cause the microspore abortion.

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Correlation between Reproductive Allocation and Elevation of Saussurea graminea in Eastern Margin of the Tibetan Plateau

MA Yi-Xin, WANG Yi-Feng, LI Yi-Ying, WANG Wen-Yue, KOU Jing

Bulletin of Botanical Research 2017, 37 (1): 23-30. DOI: 10.7525/j.issn.1673-5102.2017.01.004

Abstract （720）

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Westudied the effect of elevation on the reproductive allocation of Saussurea graminea in Qinghai-Tibetan Plateau, China.We used the field investigation method and dry mass method to evaluate morphological characteristics of S.graminea at different elevations. With the increase of altitude,the reproductive allocation,each tubula r loret mass and mass of pistil were positively correlated with elevation, but individual size, reproductive organ biomass, vegetative organ biomass, tubular loret number and mass of stamenwere negatively correlated with elevation; the reproductive allocation is negatively correlated with individual size. There are trade-offs of reproductive allocation in tubular loret number and mass as well as mass of pistil and mass of stamen. Theelevation had a significant impact on the biomass and reproductive allocation of S.graminea.The altitude could not fully explain the change of reproductive allocation strategy of this alpine perennial, and the reproductive allocation is limited by individual size of S.graminea. The characteristics of vertical distribution of S.graminea is likely to be completed by the individual size of the plant.

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Morphological Diversity of Gemma Cups on Marchantia paleacea subsp. diptera and Gemma Spread at Karst Mountains Area in Winter

SHANG Guan-Min, WANG Zhi-Hui, ZHANG Zhao-Hui

Bulletin of Botanical Research 2017, 37 (1): 31-36. DOI: 10.7525/j.issn.1673-5102.2017.01.005

Abstract （1105）

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Gemma cup is Marchantia peculiar asexual reproductive organs, but there is little study on its morphological characteristics and spread behavior in winter. With Marchantia paleacea subsp. diptera at Guizhou karst mountain area, we observed the gemma cup and gemma output in wild, and analyzed sample in the coldest winter in January 2016. The gemma cups form were varied. The gemma cups were divided into four growth period:immature period(transparent), mature period(green), recession period(purple) and decline period(atropurpureus) according to their color and gemma characteristics. The four growth periods reflected the different stages of growth and development of gemma cup. Each growth period had a different number of gemma cup, showing apoptotic characteristics orderly. In 708 gemma cups, the numbers of four period were 62, 209, 254 and 183, respectively. The average output of gemmae in different period of gemma cup was significantly different, the average gemmae output of four-period cups were 42, 131, 87 and 0, respectively. The densities of the gemma cup and gemma on the gametophyte were higher in winter, each square meter reached 10139 and 754889. Except the common by raindrops tapping spread, the winter gemma could spread by gravity. Therefore, M.paleacea subsp. diptera gemma cup in different periods and the formation of gemma are still very rich, and with the positive survival significance for the species to adapt to severe environmental conditions at karst mountains area in the cold months.

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Effects of Biological Soil Crusts on Growth and Nutrient Uptake in Three Desert Herbs in the Gurbantunggut Desert,Northwestern China

ZHUANG Wei-Wei, ZHOU Xiao-Bing, ZHANG Yuan-Ming

Bulletin of Botanical Research 2017, 37 (1): 37-44. DOI: 10.7525/j.issn.1673-5102.2017.01.006

Abstract （933）

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As an important component for desert ecosystem, biological soil crusts(biocrusts) may possibly affect adjacent vascular plants via hydrology, soil erosion and nutrient cycling, while experimental evidences are still scarce and controversial. In the present research, we attempted to experimentally investigate whether and how biocrusts affect growth and nutrient uptake in vascular plants of the Gurbantunggut Desert. We conducted the experiments to examine the effects of biocrusts on growth and nutrient uptake in three widely distributed species of Erodium oxyrrhynchum, Alyssum linifolium and Hyalea pulchella. The results showed that the effects of biocrusts on the growth of three desert vascular plants were different in different growing phases. The biomass the three species in crusted soils were higher than those in uncrusted soils in the early growth period and lower in the later part of the growing period. While biocrusts did not significantly affect the biomass allocation between shoot and root. Biocrusts also influenced nutrient uptake by plants, especially promoting uptake of N and K, while did not affect the uptake of P. The influence on uptake of other elements(Cu, Ca, Mg, Na, Cl) was species-specific. We expect to provide important scientific basis on the potential vegetation succession direction in this desert ecosystem.

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Effects of CO ₂ Enrichment on Chlorophyll Fluorescence Characteristics of Vallisneria natans

HAN Yan-Qing, LIU Xin, HU Wei-Ping, ZHANG Ping-Jiu, DENG Jian-Cai, CHENG Ze-Lin

Bulletin of Botanical Research 2017, 37 (1): 45-51. DOI: 10.7525/j.issn.1673-5102.2017.01.007

Abstract （844）

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To explore the effects of elevated atmospheric CO ₂ on photosynthetic physiology of submerged plant, we studied the effects of high concentrations of CO ₂ on chlorophyll fluorescence characteristics of Vallisneria natans by measuring the rapid fluorescence induction kinetics curves(OJIP) of the leaves with Plant Efficiency Analyzer and analyzing with JIP-test. The V _j, M _o of V.natans were significantly increased and S _m, ψ _o and φ _Eo were significantly reduced after 60 days, and the electron transfer diminished capacity of V.natans was decreased compared with the control; the donor side of PSⅡ reaction center was hurt because the Wk was increased; the specific activity parameters ABS/RC, DI _o/ RC, TR _o/ RC, DI _o/ CS _o were significantly increased, ET _o/ RC, RE _o/ RC, ET _o/ CS _o, RE _o/ CS _o significantly decreased, and the energy for heat dissipation significant increased lead to the energy of electron transfer and electron transfer chain end were significantly decreased. Compared with the control, the performance parameters F _v/ F _m and PI _abs were also decreased. Therefore, the long-term high concentrations of CO ₂ suppressed photosynthetic apparatus state, decreased PSⅡreaction center activity and down regulated photosynthetic function of V.natans, and the V.natans appeared light acclimation.

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Subcellular Localization and Expression Analysis of Genes from Eucommia ulmoides Involved in MVA and MEP Pathway

WANG Lin, DU Hong-Yan, WUYUN Ta-Na

Bulletin of Botanical Research 2017, 37 (1): 52-62. DOI: 10.7525/j.issn.1673-5102.2017.01.008

Abstract （1507）

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The full-long of EuDXR, EuMCT, EuCMK, EuMDS, EuACOT, EuHMGS and EuHMGR genes were isolated from Eucommia ulmoides. The structure characteristics were analyzed by using bioinformatics, which revealed that these deduced proteins had 73%-85% identities with the corresponding protein from other plants. Recombinant plasmid was introduced into the lower epidermal leaf explants of tobacco, and the transient expression of these genes with GFP fusion protein were observed with a Zeiss LSM. The subcellular localization analysis showed that the EuDXR, EuMCT, EuCMK, EuMDS protein were all located in the chlorophyll, EuACOT and EuHMGR protein were both located in the endoplasmic reticulum, EuHMGS protein was located in the plasma membrane. By RNA sequencing, the spatial-temporal expression patterns of these genes were detected. The expression of genes in MEP pathway is higher in leaves, but the expression of genes in MVA pathway is higher in young fruit. The rubber content in young fruit was higher than that in leaves. Therefore, the MVA pathway played a dominant role in the biosynthesis of Eu-rubber.

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Expression and Stress Tolerance Charaterization of a ThDREB Gene from Tamarix hispida in Yeast

FENG De-Ming, WEN Pei-Ying, ZHAO Chang, YANG Yuan-Biao, YANG Gui-Yan, YU Li-Li, GAO Cai-Qiu

Bulletin of Botanical Research 2017, 37 (1): 63-68. DOI: 10.7525/j.issn.1673-5102.2017.01.009

Abstract （833）

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DREB protein could specifically bind to DRE(dehydration responsive element) cis-acting elements, and regulate the expression of downstream stress related gene. It plays an important role in the regulation responses of plants to abiotic stress. To explore the abiotic stress tolerance role of DREB(ThDREB) from Tamarix hispida, the recombinant plasmid(named as pYES2-ThDREB) was constructed by inserting the ThDREB gene into the yeast expression vector pYES2. The plasmid pYES2-ThDREB was transformed into the yeast Saccharomyces cerevisiae, and the yeast transformed with empty pYES2 was used as the control. The pYES2-ThDREB and pYES2 recombinant yeast were treated with sorbitol, H ₂O ₂, CdCl ₂, NaCl, Na ₂CO ₃, MgCl ₂, and -20℃, respectively. The pYES2-ThDREB recombinant yeast was tolerant to all above abiotic treatments, suggesting that ThDREB maybe an important stress-regulating genes in T.hispida.

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Sequence and Expression Analysis of MADS-box Genes in Celastrus orbiculatus Thunb. Based on RNA-seq Data

ZU Kui-Ling, DONG Shu-Bin, LI Jian-Xia, ZHAO Yun-Yu, ZHAO Liang-Cheng

Bulletin of Botanical Research 2017, 37 (1): 69-77. DOI: 10.7525/j.issn.1673-5102.2017.01.010

Abstract （895）

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For the regulatory information of MADS-box genes on the flower and fruit development in Celastrus orbiculatus Thunb, 15 MADS-box genes( ColMADS) with full ORF(Open Reading Frame)were identified from transcriptome generated from female flower and fruit. The features of the proteins were analyzed by the bioinformatics methods. The comp37814_g1and comp41380_g2 belonged to M-type family, while other 13 genes were the members of MIKC. All were unstable hydrophilic proteins except for comp37814_g1. Alpha helix, beta turn, extended strand and random coil were found from the secondary structure of ColMADS protein, and alpha helix was major part. Five motifs were identified. The Motif 1 and Motif 2, both consisting of 50 amino acids, were the conserved structure of MADS-box family and only Motif 1 contained MADS domain. By phylogenetic analysis, 15 ColMADS genes were divided into 10 clades, belonging to different subfamilies and groups. In addition, the expression patterns of the genes among different development stages were analyzed using real-time RT-PCR. Two genes were highly expressed at flower blooming stage and nine genes were obviously expressed at flower senescence stage. There are four genes upregulated during the young fruit development.

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Evaluation,Selection and Application of Dendrobium officinale EST-SSR Primers

HU Zhong-Yi, FU Tao, HE Yue-Qiu, LI Wen, LIN Li

Bulletin of Botanical Research 2017, 37 (1): 78-87. DOI: 10.7525/j.issn.1673-5102.2017.01.011

Abstract （890）

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By developing and evaluating EST-SSR primers, we studied their application in genetic diversity, species identification, genetic map construction, genetic relationship identification of Dendrobium officinale. The early developed 20 pairs of primers with good polymorphism were used for the analysis of 16 samples(15 samples of D.officinale and 1 sample of D.falconeri) by polyacrylamide gel electrophoresis, and the amplified bands were calculated. Nei's gene diversity and Simpson index were tested using PopGen32 to analysis the differentiation rate of single primer and combination primers. Finally, the construction of genetic similarity coefficient clustering map was calculated by using NTSYSpc2.1. The 16 pairs of primers, such as DN4, were highly polymorphic, while lower in DN13, DN23, DN60 and DN67. Although single primer could not distinguish all the samples, DN4, DN10, DN105, DN81, DN39 and DN71 displayed relatively higher distinguish rate. All the samples could be distinguished effectively by combined primers, and the DN4+DN10+DN105+DN39 primer set could be used as the core primer for fingerprint construction and genetic diversity analysis of D.officinale. By cluster analysis, the 20 pairs of primers could be used for the identification of both intraspecific and interspecific phylogenetic relationships of D.officinale. The primers DN13, DN39, DN4, DN58, DN65, DN67, DN10 and DN99 could be used for the identification of D.officinale and D.falconeri. The 20 pairs of primers could be wildly used in the application of genetic diversity analysis, germplasm identification, genetic relationship identification and genetic map construction of D.officinale.

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Assessment of Genetic Diversity of Rhododendron nivale in Tibetan, China using AFLP Markers

XU Jing-Jing, ZHAO Bing, ZHANG Liang-Ying, SHEN Hui-Fei, LI Hou-Hua

Bulletin of Botanical Research 2017, 37 (1): 88-95. DOI: 10.7525/j.issn.1673-5102.2017.01.012

Abstract （841）

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To determine the correlation among genetic variations, the geographic location of a population, and factors that influence high-level genetic diversity, the genetic diversity of 135 Rhododendron nivale samples was studied using amplified fragment length polymorphisms(AFLP). A total of 273 amplification products were generated from 6 selective primer combinations, of which 85.71% were polymorphic. The results of POPGENE showed the change trends of the percentage of polymorphic( PPL), Nei's genetic diversity( h) and Shannon's index(I) were similar. The change trends of Nei's genetic diversity( h) and Shannon's index(I) were Gongbujiada < Milin < Galongla < Sejila Mountain < Hongla Mountain. The POPGENE analysis showed that R.nivale had higher genetic diversity at species level( PPL=85.71%, I=0.415 1, h=0.273) but lower at population level( PPL=62.26%, I=0.280 3, h=0.184 1). Both the results of POPGENE( G _st=0.324) and AMOVA(36% among populations, 64% within populations) showed that the most genetic variance existed within the populations. UPGMA tree plots indicated that there was no significant correlation between the genetic distance and the geographic distance among the R.nivale populations. At last, some conservation strategies for R.nivale germplasm were put forward.

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Effects of Different Substrate Materials on Growth Biomass and Nutrient Element Contents of Camellia oleifera Container Seedlings

CUI Na-Na, ZHAN Chang-Sheng, HU Juan-Juan, CAO Zhi-Hua, SHU Qing-Long

Bulletin of Botanical Research 2017, 37 (1): 96-103. DOI: 10.7525/j.issn.1673-5102.2017.01.013

Abstract （819）

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The experiment was conducted to study the effects of different substrate material on Camellia oleifera container seedlings, and find out the local substrates instead of peat soil content. One-year-old C.oleifera container seedlings of different substrate treatments were tested in several indicators such as survival rate, growth, biomass, the contents of chlorophyll and nutrient elements aboveground, and the correlation between substrate and contents of nutrient element aboveground was also analyzed. The indexes including survival rate, height of seeding, ground diameter, ratio of height to ground diameter, tap root length, tap root/lateral root fresh weigh, aboveground/underground biomass, the diameter root ratio/FW, the diameter root ratio/DW, and contents of chlorophyll were superior in treatment Ⅰ(50% Peat soil+50% the rest of the same composition) and treatment Ⅴ(5% the rest of edible mushroom+5% fowl dung+5% cow dung+35% Peat soil+50% the rest of the same composition), and there was significant difference between them and other three treatments. There was significant difference in the contents of N, P, K, Ca, S, Fe, Mn, Cu and Zn, and no significant difference in B and Mg between different treatments. The correlation analysis showed that Mg and available potassium, organic matter had significantly positive correlation, and others had negative correlation. Therefore, the best substrate was treatment Ⅴ, in which local substrate instead of part of the peat soil. The study would provide a new way of reducing seedling costs and maintaining sustainable utilization of substrate.

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Effects of Charcoal on Seed Germination of Larix gmelinii

CAO Feng-Yan, MAO Zi-Jun, QU Lai-Ye

Bulletin of Botanical Research 2017, 37 (1): 104-108. DOI: 10.7525/j.issn.1673-5102.2017.01.014

Abstract （804）

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To understand the effects of charcoal after forest fire on the germination of Larix gmelinii at Great Xingan Moutains, we investigated the charcoal distribution and collected different types of charcoal produced by trees( L.gmelinii and Betula platyphylla Suk.), shrub and grass, respectively. We collected charcoals produced by different years(1, 2, 4 and 5 year) from the burned forest sites. Then, the charcoals were soaked into water at 12, 24, 48 and 96 h to obtain different charcoal solution. There were significant effects of different charcoal and different produced years on the germination of L.gmelinii seeds. However, the strength and directions of effects were randomly, and the effects of charcoal were very completed.

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Basic Characteristics of Soil Seed Bank of Haloxylon ammodendron in Junggar Basin

LÜ Chao-Yan, ZHANG Xi-Ming, GAO Zhi-Xi, XIONG Bai-Lian, HAN Chang, WANG Jia-Zhen

Bulletin of Botanical Research 2017, 37 (1): 109-117. DOI: 10.7525/j.issn.1673-5102.2017.01.015

Abstract （926）

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Soil seed bank is the refuge of seed and the basis of vegetation regeneration and recovery in arid desert region. Haloxylon ammodendron-based desert vegetation is the important ecological barrier of Junggar Basin's oasis area, however, the researches of soil seed bank characteristics which are closely related to population regeneration of H.ammodendron are relatively lacking. Basic characteristics of the soil seed bank of H.ammodendron in Junggar Basin were researched by field sampling,physical screening and laboratory experimenting. The results showed that:(1)The soil seed bank densities of H.ammodendron populations varied from 71 to 696 seed·m ^-2, especially in some local environments it reached 7 534 seed·m ^-2. (2)The coefficient of variation analysis identified that the coefficients of variation of the soil seed bank density were more than 1. It meant that the quantitative distribution of soil seed bank was very uneven. (3)The seed germination rates in the soil seed banks varied from 2.08% to 47.62% and the average was approximately 18%. (4)The analysis of variogram indicated that there were obvious differences in the spatial distribution of seeds in the soil seed bank of H.ammodendron populations. In summary, the characteristics of high density, germination ability and spatial variability would effect on the distribution pattern and regeneration of H.ammodendron populations.

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Sensitivity of Fruiting for Pinus koraiensis to Climate Change and Mechanisms of Masting in the Original Broad-leaved Korean Pine Forest in North Xiaoxing'an Mountain,China

CHENG Chun-Xiang, MAO Zi-Jun, JIN Shi-Bo, SONG Guo-Hua, SUN Peng-Fei, SUN Tao

Bulletin of Botanical Research 2017, 37 (1): 118-127. DOI: 10.7525/j.issn.1673-5102.2017.01.016

Abstract （936）

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To find out sensitivity of fruiting for Korean pine( Pinus koraiensis) to climate change and mechanisms of masting, we studied Korean pine in the original broad-leaved Korean pine forest which has high stand stability and the yields of Korean pine seeds in nearly 16 years. We decomposed the yields of Korean pine seeds into trend yields and climatic fluctuant yields by resouce switching theory, resource matching theory and the suggestion of Nicholls. Then, we analyzed the influence of weather on climatic fluctuant yields. The results reveal:(1)The current year yields of Korean pine seeds significantly negative correlated with the combined yields of the last 2 years and showed exponential relationship with it, which supports the resource switching hypothesis. And resource switching probably acts as Korean pine cone maturating inhibition among the reproductive processes in a mast year. (2)The temperature difference between the two previous Junes(ΔT ₆) better predicts the yields of Korean pine seeds. (3)The period of bud meristerms development for Korean pine fruiting was the most key climate-sensitive phase. Negative temperature and sunshine duration, coupled with positive precipitation in the period significantly correlated with climatic fluctuant yields of Korean pine. (4)Positive temperature anomaly in some years was beyond the optimum temperature range of the development of bud meristerms and caused reproduction of Korean pine to fall. (5)The yields of Korean pine seeds in many periods of reproductive development was sensitive to climate change, indicating that resource switching and resource matching may be exhibited by a single masting. These findings suggest Korean pine reproduction are vulnerability to climate warms and may be an increasingly important bottleneck for future Korean pine regeneration, especially in areas with greater increases in temperature.

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Dynamics of Leaf Litter and Mineral Soil Nutrients in Different Sized Pinus massoniana Forest Gaps

WEI Da-Ping, ZHANG Dan-Ju, LI Xun, ZHANG Yan, LI Chuan-Bei, ZHANG Jian

Bulletin of Botanical Research 2017, 37 (1): 128-138. DOI: 10.7525/j.issn.1673-5102.2017.01.017

Abstract （904）

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In order to study the organic carbon nitrogen phosphorus nutrients content, loss of these nutrients, C/N/P stoichiometric ratio of Pinus massoniana litter and soil in different sized P.massoniana forest gaps. In the present study, with existing 42 years of P.massoniana plantation, by cutting, formed four different sized P.massoniana forest gaps about 100, 400, 900, 1 600 m ² and without cutting as usual. Bying leaf litter decomposition method. The results showed that:(1) With different sized forest gaps, the organic carbon nitrogen phosphorus nutrients content, loss of these nutrients and C/N/P stoichiometric ratio of P.massoniana litter and soil had significant differences, in addition to the phosphorus nutrients content of soil and phosphorus nutrient loss. With the increasing of gap size G1~G4, the content organic carbon nitrogen phosphorus nutrients content of P.massoniana litter all showed a decreasing trend and had smaller values in G3. Most index of the content organic carbon nitrogen phosphorus nutrients content of soil, the content organic carbon nitrogen phosphorus nutrient loss showed a parabolic trend and had the maximum value in G2 or G3; (2) With different sized forest gaps, the C/N/P stoichiometric ratio of P.massoniana litter and soil had significant differences. With the increasing of gap size G1~G4, the C/N/P stoichiometric ratio of soil showed a parabolic trend, C/N had the maximum value in G3, both C/P and N/P had the maximum value in G2. The Soil C/N, C/P and N/P coefficient of variation were 13.31%, 16.51% and 17.21% respectively. Both C/N and C/P of P.massoniana litter had the minimum value in G3; (3) There were a strong correlation relationship between the organic carbon nitrogen nutrients content of P.massoniana litter and the organic carbon nutrients content, C/N/P stoichiometric ratio of soil, environmental factors, while a weaker correlation relationship between phosphorus nutrients content of P.massoniana litter and them. There were a strong correlation relationship between C/N of P.massoniana litter and the phosphorus nutrients content, C/N/P stoichiometric ratio of soil, environmental factors. There were a strong correlation relationship between C/P, N/P of P.massoniana litter and the C/P of soil, environmental factors. There were a strong correlation relationship between organic carbon nitrogen nutrient loss and the organic carbon C/N, C/P of soil, environmental factors. There were a weaker correlation relationship between phosphorus nutrient loss and the organic carbon nitrogen phosphorus nutrients content, C:N:P stoichiometric ratio of soil, environmental factors. There were a strong correlation relationship between the organic carbon nitrogen phosphorus nutrients content, C/N/P stoichiometric ratio of soil and environmental factors.

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Growth Traits Variation and Early Selection of Mytilaria laosensis Provenances and Families

QIN Min, YIN Guang-Tian, YANG Jin-Chang, LI Rong-Sheng, ZOU Wen-Tao

Bulletin of Botanical Research 2017, 37 (1): 139-146. DOI: 10.7525/j.issn.1673-5102.2017.01.018

Abstract （761）

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We selected the fast growing provenance-families of Mytilari laosensis to analyze the variation regularity of growth traits among provenances and among families of M.laosensis. We chose 116 families within 8 provenances of M.laosensis, and used genetic variance analysis and breeding value to analyze the growth traits among provenances and among families of M.laosensis. There were highly significant differences among growth characters at family and provenance level. At the age of 6, the average of height(H), diameter at breast height(D) and volume(V) of provenance GXPX was 9.50 m, 13.40 cm and 0.079 9 m ³, and that was 1.28, 1.42 and 2.30 times of worst provenance GXFC, respectively. The average of H, D and V of family FC01 was 9.86 m, 15.31 cm, 0.1 016 m ³, respectively. Mean individual volume of family PX01 was 4.01 times of worst family FC01 and 1.94 times of the average of total families by comparison. Using individual trait selection, two superior provenances were selected from eight provenances population tested, and genetic gain of H, D and V of those superior provenances reached as high as 8.87%, 11.79% and 34.70%, respectively. By using grade of breeding value selection, 12 superior families were selected from 123 families population tested, and genetic gain of H, D and V of those superior families reached as high as 11.57%, 17.35% and 52.06%, respectively. At the age of 6, the comprehensive growth of the best excellent provenances were GXPX and GXLZ, and the comprehensive growth of the best excellent families were PX01, LZ03, PX02, DB08, PX10, PX04, PX09, DB07, DB16, LZ05, JX16 and PX05. These superior provenances and families can be used as seed material to declare and promote.

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Comparative Study on Staining Methods and Techniques of Cell Wall Histochemistry

ZHANG Xia, HU Lu-Jie, ZHOU Cun-Yu, YANG Chao-Dong

Bulletin of Botanical Research 2017, 37 (1): 147-154. DOI: 10.7525/j.issn.1673-5102.2017.01.019

Abstract （2636）

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We carried out the comparative studies on several methods and techniques for cell wall histochemistry under light and epifluorescence microscope. The results showed that:(1) To study Casparian bands using staining methods of sulphuric acid digestion and berberine hemisulfate-aniline, the requirements of material collecting time and part were high, therefore, it was recommended to use the two methods to mutual confirmation; (2) To study suberin lamellae using Sudan red 7B staining, and none staining and berberine hemisulfate-aniline blue staining under blue excited, none staining was more sensitive than Sudan red 7B, but Sudan red 7B staining is convenient under light microscope; (3) To study lignified walls, berberine hemisulfate-aniline staining was more sensitive than phloroglucinol-HCl; (4) Using TBO fast wall staining to replace Sudan Ⅲ/Ⅳ conventional staining, the wall borders and layers were clearer.

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Effect of Cu on Compositions of Seed Oil in Paeonia ostii

ZHAO Xiao-Ju, QIN Wei, CHEN Hua-Feng

Bulletin of Botanical Research 2017, 37 (1): 155-160. DOI: 10.7525/j.issn.1673-5102.2017.01.020

Abstract （774）

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The unsaturated fatty acids, especially linolenic acid are the main embodiment of the quality of peony seed oil. But so far, in traditional cultivation area of Paeonia ostii, it has not been investigated whether the soil copper content affected the quality of peony seed oil. In this study, we investigated copper content in soil and the composition of the seed oil in 30 P.ostii cultivation areas in Anhui Province. The copper content in the soil was 18.98-298.82 mg·kg ^-1, and the variation coefficient was 83.06%. The contents of palm acid, stearic acid, oleic acid, linoleic acid and linoleic acid were 5.62%, 1.89%, 24.59%, 29.76% and 38.13%, respectively. The variation coefficient was between 5.66 and 9.72, among which the highest one was linoleic acid. There was a significantly negative correlation between copper content and unsaturated fatty acid content, linoleic acid content, and there was no significant correlation between the content of copper and linolenic acid content. And there was a significantly positive correlation between soil copper content and leaf copper content, r=0.778. The cultivation of P.ostii for oil production should avoid the high copper content in the soil to keep the quality of peony seed oil.

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Correction of A Typographical Error in the Protologue of Clematis chaohuensis

LIN Yun, SHAO Qing, YANG Zhi-Rong

Bulletin of Botanical Research 2017, 37 (2): 161-163. DOI: 10.7525/j.issn.1673-5102.2017.02.001

Abstract （1445）

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In the course of the normalized compilation of type specimens which deposited at China National Herbarium(PE), Institute of Botany, Chinese Academy of Sciences, we discovered an error in the citation of type collection of Clematis chaohuensis W.T.Wang & L.Q.Huang(Ranunculaceae). The holotype of C.chaohuensis W.T.Wang & L.Q.Huang was incorrectly cited as D.Q.Wang 70543 in the protologue, but is actually D.Q.Wang 95043.

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Geographical Distribution Pattern of Macromitrium in the World and Its Biogeographical Significance

GUO Shui-Liang, WU Qian-Qian, YU Jing, CAO Tong

Bulletin of Botanical Research 2017, 37 (2): 164-173. DOI: 10.7525/j.issn.1673-5102.2017.02.002

Abstract （1334）

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The genus Macromitrium(Orthotrichaceae, Bryophyta) is an important taxon in bryological systematics, ecology and biological conservation. On the basis of our taxonomical revision of the genus, we got the distribution data of 274 species of the genus in 14 geographical unites(suggested by "Index Muscorum") in the world.By using Clustering Analysis and Detrended Correspondence Analysis(DCA), we got a dendrogram and a three-dimensional scatter plots to reveal the distribution pattern of the genus visually. The results showed a distinctive floristic differentiation of Macromitrium among As2, Oc, As3-4, Afr2-3-4, Am2-3-4-5, and Austr1-2, with highest diversity in Tropical Asia and Tropical America. Except M.microstomum(Hook.& Grev.) Schwägr., all other species were recorded from a single or a few geographical unites. Among different global regions, Macromitrium differentiated not only in its floristic elements, but also in their morphological characters. We discussed the systematic and biogeographical significances of the genus according its global distribution pattern.

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Reduction on Orinus longiglumis with Generalized Morphological Characters and nrDNA ITS Sequence

LIU Yu-Ping, LÜ Ting, ZHOU Yong-Hui, ZHANG Xiao-Yu, SU Xu

Bulletin of Botanical Research 2017, 37 (2): 174-180. DOI: 10.7525/j.issn.1673-5102.2017.02.003

Abstract （1696）

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Orinus is an alpine genus with important economic values in the grass family(Poaceae). There are six species in it and all of them have stronger adverse-resistant characteristics, and they are the important resources for improving and applying agricultural breeding and animal husbandry. Meanwhile, the discriminative characters are obscure among these species. We compared and analyzed the main morphological characters and molecular sequence differences among nearest related Orinus longiglumis, O.thoroldii and O.anomala by generalized morphological observation and nuclear ITS region research. There is very slight disparity between O.longiglumis and O.thoroldii in generalized morphological characters, the classification of the two species is very difficult, the differences between O.longiglumis and O.anomala are distinct, and the two taxa are two different species. We further used the ITS sequence variation to tentatively delimit the previously acknowledged species and found that there is no distinct boundary between species and only two distinct lineages were recognized for all three species. A branch is composed of O.longiglumis and O.thoroldii, while another is only made up of O.anomala. The bootstrap support rates of two branches reach as high as 100%. Therefore, O.longiglumis and O.thoroldii should belong to the same species, and O.longiglumis can be reduced to synonymy of O.thoroldii.

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Microsporogenesis,Male Gametogenesis and Pollen Morphology of Gonocaryum lobbianum

KONG Dong-Rui, LI Lu

Bulletin of Botanical Research 2017, 37 (2): 181-184. DOI: 10.7525/j.issn.1673-5102.2017.02.004

Abstract （1441）

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We studied the microsporogenesis, male gametogenesis and pollen morphology of Gonocaryum lobbianum using light microscopy and scanning electron microscopy to enrich the embryological and palynological data of Cardiopteridaceae s.l. The main results are as follows:(1) The anther is tetrasporangiate; (2) The anther wall comprises the epidermis, fibrous endothecium, ephemeral middle layer and secretory tapetum with 2-4 nuclei in each cell; (3) The microspore mother cell undergo meiosis, cytokinesis is simultaneous and the tetrads are tetrahedral; (4) The mature pollen grains are two-celled; (5) The pollen grains are cryptoaperturate with three pores and the extin is reticulate. The characters of microsporogenesis and male gametogenesis of Gonocaryum are similar to those of Cardiopteris with a little difference. G.lobbianum is almost the same as the other species in the pollen morphological characters.

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The Research of Key Influence Factors of Rapid Culture of the Protonema of Didymodon ditrichoides

ZHAO Yang, LI Xiao-Ming, LI Ru-Xue, WANG Chun, BU Chong-Feng

Bulletin of Botanical Research 2017, 37 (2): 185-193. DOI: 10.7525/j.issn.1673-5102.2017.02.005

Abstract （1348）

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In order to provide a rich source of mass vaccination for the field restoration of moss crust, we used two kinds of disinfection method, 0.1% NaClO(10, 15, 20, 30, 45, 60, 120 s) and 0.1% HgCl ₂(10, 15, 20, 30, 45, 60, 120 s), with Knop, MS and Hoagland as culture media, and set the pH values(5.5, 6, 6.5, 7, 7.5, 8, 8.5), through the single factor experiment design method, we determined the survival rate of the cormus of[ Didymodon ditrichoides(Broth.)], protonema length and number of branches, thereby exploring key factors that affect the propagation of the protonema of D.ditrichoides. The results are as follows:(1) The disinfection method, culture medium and pH value have significant influence upon the survival rate of the cormus of D.ditrichoides, length of protonema and number of branches( P<0.05), in a descending order of disinfection method, culture medium, and pH value; (2) The most suitable disinfection method is 0.1% NaClO with a duration of 15-20 s; (3) The most suitable culture medium for the growth of the protonema of D.ditrichoides is Knop and Hoagland; (4) The most suitable pH value for the growth of protonema of D.ditrichoides is 7.5. Therefore, the best combination of factors for rapid cultivation of the protonema of D.ditrichoides is 0.1% NaClO disinfection 15-20 s+Hoagland/Knop+pH=7.5. Our experiment can be determined to shorten the growth cycle of D.ditrichoides and realize rapid propagation, thereby providing experience for the rapid culture and engineering application of moss crust.

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Construction and Expression of mCherry Expression Vector Used in Plant

YU Ying, LIU Jia-Xin, ZHOU Mei-Qi, ZHAO Lei-Fei, WANG Chao

Bulletin of Botanical Research 2017, 37 (2): 194-199. DOI: 10.7525/j.issn.1673-5102.2017.02.006

Abstract （2966）

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Fluorescent proteins were widely used and played important roles in the biological research. The red fluorescent protein mCherry is valuable to the plant research because of its color and stability. The mCherry gene was inserted into pBI121 plant expression vector, and the pBI121MCS-mCherry vector was constructed. The vector were transferred into onion epidermis by particle bombardment, and the whole onion cells showed red fluorescence under the microscope, which suggested that the vector could express red fluorescent protein in plant cells. The transcription factor BpMYB4 gene was constructed in this vector using double enzyme digestion, and the fusion expression vector pBI121MCS-mCherry-BpMYB4 was obtained and expressed in onion epidermis. Only the nucleus showed red fluorescence, which indicated that the vector can be used to express the fusion protein and used in the subcellular localization accurately. While the fusion expression vector can be constructed simply, and the intermediate vectors are no longer required. The KpnⅠ sites was introduced which can help the fusion of more genes. Therefore, mCherry vector can be used in cell localization of plant genes and screening of transgenic plants. This study would provide materials for genomics research of birch in future.

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Research on Understory Vegetation and Soil Physical-chemical Properties of Teak Plantation with Difference Stand Densities

ZHOU Shu-Ping, LIANG Kun-Nan, DU Jiang, LI Bi-Jun, ZHOU Zai-Zhi, HUANG Gui-Hua

Bulletin of Botanical Research 2017, 37 (2): 200-210. DOI: 10.7525/j.issn.1673-5102.2017.02.007

Abstract （1008）

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We studied the effect of teak( Tectona grandis L.f) plantations with different densities on understory vegetation and soil physical-chemical properties for providing theoretical basis for establishment and sustainable management of teak plantations. We used teak plantations at age of 14-16 with different stand densities(650, 900, 1 050, 1 200 and 1 450 tree·hm ^-2) in Jieyang City of Guangdong Province in the experiment. We recorded the species name, quantity, height and coverage of understory vegetation by plant quadrate, and measured soil physical-chemical properties by collecting soil samples from 0-20 and 20-40 cm depths, respectively. We investigated the diversity index of understory vegetation and soil physical-chemical property to evaluate site quality of teak plantation under different densities by principal component analysis. The coverage of understory vegetation was decreased with the increase of stand density. The dominant species of herbs were changed from sun species to shade-tolerant species with the increase of stand density, and the Shannon-Wiener index, Simpson index, Richness index, and Evenness index jointly showed a tendency that falls after increases first along with the raise of stand density. The physical-chemical properties in the upper layer of soil were superior to these in the deeper layer of soil at the same density. With the increase of stand density, the soil physical-chemical properties presented a change process from first improvement to degradation. The capillary porosity, non-capillary porosity, available K, available P, total P, exchangeable acid and exchangeable aluminum of soil showed significant difference among the different stand densities( P<0.05). By biodiversity indexes and physical-chemical properties of principal component analysis, the order of the composite score of different stand densities on species diversity and soil physical-chemical properties from high to low was:1 050 tree·hm ^-2(4.82), 900 tree·hm ^-2(1.58), 650 tree·hm ^-2(-1.30), 1 200 tree·hm ^-2(-1.81) and 1 450 tree·hm ^-2(-3.29). By principal component analysis, the stand density(1 050 tree·hm ^-2) was more favorable to maintaining better undergrowth species diversity and soil physical-chemical properties. Stand density would be adjusted reasonably according to the actual situation in the management process of teak plantation.

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Expression Analysis of Rev1 and Rev3 of Halostachys caspica under Salt Stress

DU Chi, ZHANG Ji, ZHANG Fu-Chun

Bulletin of Botanical Research 2017, 37 (2): 211-215. DOI: 10.7525/j.issn.1673-5102.2017.02.008

Abstract （978）

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According to the transcriptome of Halostachys caspica under salt stress, the real-time PCR primers designed about Rev1 and Rev3 were referred to the ESTs of transcriptome sequencing. We established a method for detecting of mRNA transcription of Revs by Real-time PCR, and the transcription level of Revs in different concentration of NaCl present. Two genes have similar expression pattern, the expression levels remain stable under 100 mmol·L ^-1 NaCl treatment. As the concentration of NaCl was increased and the stress time was prolonged, the genes relative expression levels were also gradually increased. HcRev1 gene in 700 mmol·L ^-1 NaCl stress for 14 d was rapidly up-regulated and reached the peak level, which was 4.63-fold of the control. HcRev3 gene in 300 mmol·L ^-1 NaCl stress for 14 d was 15.55-fold of the control with significant differences. Both HcRev1 and HcRev3 genes were induced by salt stress, although there were different expression levels of HcRev1 and HcRev3 genes, which were involved in the DNA damage repair during salt stress.Our study could be used to clarify the regulatory function of HcRev1 and HcRev3 genes between DNA damage-repair and salt resistance in plants.

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C,N and P Stoichiometry at Different Stages of Litter Decomposition in Subalpine Forest of Western Sichuan Province and Interspecies Comparison

SHEN Xian, LIU Yang, DENG Jing, SHI Jia-Qi

Bulletin of Botanical Research 2017, 37 (2): 216-226. DOI: 10.7525/j.issn.1673-5102.2017.02.009

Abstract （3076）

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We used spatial sequence instead of time successional sequence to study the stoichiometry of litter decomposition in subalpine forest of Western Sichuan. Three litter layers including Fresh litter layer(L), Fermentative layer(F) and Humus layer(H) under natural conditions were taken to simulate different litter decomposition stages, studied on Carbon(C), nitrogen(N), phosphorus(P) and dissolved organic carbon(DOC), soluble nitrogen(SN) and phosphorus(SP) contents of three litter layers of Picea asperata(Pa), Abies faxoniana(Af), Betula platyphylla Suk( Bp) and Rhododendron lapponicum(Rl) in four subalpine forests. The forest type and decomposition stages significantly affected C, N, P contents and its stoichiometric ratio in the process of litter decomposition. For all the forest litter, DOC and SP contents reduced and leached quickly at the early decomposition stage whereas slowly in the later periods of decomposition. SN, SP and DOC contents of hardwood species were generally higher than those of conifer species, which in the order of Bp > Rl > Af > Pa, especially at the early decomposition stages. There was significant difference between conifer species and hardwood species, and SN content of Pa and Af reduced with the ongoing of decomposition process while Bp and Rl got highest in the F layer. C content reduced with the ongoing of decomposition process, and N content of Af, Bp and Rl got highest when they were in the Fermentative layer(F > L > H). Litter decomposition stages had no significant effect on P content of Pa and Bp. Overall, litter C/P and N/P of subalpine forest were significantly lower than the global average, C/N, C/P, N/P, DOC/C, SN/N, SP/P also reduced with the ongoing of decomposition process. For DOC/C, Bp and Rl significant decreased at early decomposition stages, and Af significantly decreased at late decomposition stages. N:P of Af increased at first and then decreased, but for Rl showed a trend of rising in the decomposition. These results provided a comprehensive understanding of the litter decomposition process and nutrient cycling in the subalpine forest.

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Morphological Variations, Genetics and Geographic Background of Echinochloa crus-galli in China

ZOU Man-Yu, LU Yong-Liang, YIN Li-Ping, GUO Shui-Liang

Bulletin of Botanical Research 2017, 37 (2): 227-235. DOI: 10.7525/j.issn.1673-5102.2017.02.010

Abstract （1061）

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Thirty-three seeds of Echinochloa crus-galli(L.) P. Beauv. from paddy regions of nine provinces(Autonomous Region) of China were sampled and sown in the same environment. Their offspring were obtained. Sixteen morphological traits of their offspring were measured. Nine ISSR primers with good repeatability were selected and 109 ISSR loci were amplified from thirty-three E.crus-galli samples. Based on morphological traits and locus information, cluster analysis and principal component analysis on these thirty-three populations were conducted. The main results are as follows:(1)Four morphological groups were identified from these populations, among them are E.crus-galli var. zelayensis(Kunth) Farw., E.crus-galli var. mitis(Pursh) Peterm. and E.crus-galli var. praticola Ohwi; (2)Six groups were also identified from these samples by using cluster analysis based on ISSR locus data, which corresponds well with the morphological groups, especially for E.crus-galli var. praticola; (3) Obvious genetic differentiation existed among these thirty-three populations, which is obviously influenced by the geographical locations( r=0.684, n=33, P<0.001). Their morphological variation, however, is weakly affected by the geographical locations( r=0.245, n=33, P<0.2); (4) Both the morphological data and the ISSR data separated the populations of E.crus-galli var. praticola from the other populations. Therefore, the variety was suggested to elevate to species level: Echinochloa praticola(Ohwi) Guo S L, Lu Y L, Yin L P & Zou M Y.

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Flowering Biological Characteristics of Olea europaea(Chenggu 32) in the North Subtropical of China

WU Wen-Jun, LU Juan, ZHAO Meng-Jiong, SHI Yan-Hu, JIANG Cheng-Ying

Bulletin of Botanical Research 2017, 37 (2): 236-241. DOI: 10.7525/j.issn.1673-5102.2017.02.011

Abstract （1093）

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We studied the flowering phenology, sexual system and flowering characters of Olea europaea L.(Chenggu 32) grown in the north subtropical areas of China to explore the characteristics of its sexual reproduction system. We analyzed the flowering biology including the florescence process, flower shape, the position and quantity of male and hermaphrodite flower as well as the pollen morphology of chenggu32 located in the southern slope of West Qinling region. These results indicated that:(1) O.europaea L. flowering phase is concentrated in May under the climatic conditions of tropical inland North. Plant flowering individuals last 15-20 days, while full-blossom period within 4-6 days; (2) O.europaea L. has an andromonoecy sexual system and the flower organ is small. Both the male flower and hermaphrodite flower have normal developing stamens. Based on the morphological observation conventional paraffin sectioning method, the pistil of hermaphrodite flower grow normally, while the pistil of male flowers has the smaller ovary and abortive stigma; (3) The pistil is the major organ for influencing biomass between the two flower types. Compared with male flowers, biomass of hermaphrodite flower is significantly larger than male flower( P<0.01), which needs fewer resources during tree differentiation; (4)There is no significant difference of pollen amount and pollen grains between male and hermaphrodite flower. The hermaphrodite flowers developed on the top of rachis, while male flowers mainly distributed at the base and center of rachis at the ratio of 21.13% and 30.77% respectively. Therefore, Chenggu 32 has typical andromonoecy. As for male flowers of Chenggu32, the location of inflorescence has no advantage for his male function. It also has no superiority about the pollen grain quantity and vitality of male flowers. However, the appearance of male flowers increased the pollen quantity and P/O, and improved the male fitness. It reduced the plant resource waste for the reason of hermaphrodite flowers fallen, thus guaranteeing the maximum breeding in limited environmental resources.

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Effects of Different Culture Medium on Moringa Seedling Growth and Correlation analysis in Northern China

XU Hai-Jun, CHENG Xin-Yu, WANG Xiao-Fei, SUN Yu-Feng

Bulletin of Botanical Research 2017, 37 (2): 242-248. DOI: 10.7525/j.issn.1673-5102.2017.02.012

Abstract （829）

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We investigated the effects of different cultivation medium on seedling growth of Moringa oleifera Lam., PKM1 and PKM2 under the facilities in northern China, the plant height, base diameter, anatomical shape character, and analyzed relationship between cultivation medium and growth of Moringa seedling. The results showed that peat+sand+rotten mushrooms base material(3:1:1, T ₂) was better fit for Moringa seedling growth than the other three treatments which are perlite+garden soil+humus(1:2:1, T ₁), garden soil+humus+peat+sand(2:1:1:1, T ₃) and garden soil(CK). The contents of organic matter, porosity, and unit weight in T ₂ substrate are 49.97 g·kg ^-1, 76.84%, and 0.58 g·cm ^-3, respectively. The available N, P, K in T ₂ are significantly higher than other treatments( P<0.05). The growth speed of Moringa seedling in T ₂ substrate are significantly faster than other treatments. After 71-day planting, the plant height of M.oleifera, PKM1 and PKM2 are 80.2, 115.67, and 81.0 cm, respectively, increased by 38.3%, 25.7%, and 28.6% more than T ₄(CK), respectively, and the base diameter are 13.27, 17.47, and 15.08 mm, respectively, increased by 32.0%, 14.6%, and 39.0% more than T ₄(CK), respectively. The aboveground part of seedling grows rapidly under T ₂ treatment. After 60-day culture, the skin of seedling under T ₂ is completely dead and broken up. The phloem fiber and xylem are the most developed among four treatments. So relationship between eight vector factors of substrate and plant height, base diameter were analyzed, the result indicate that the cultivation medium which contain high content of organic matter, with capacity of permeability and water retention is more suitable for the growth of Moringa seedling. The factors which are significantly correlative to growth of Moringa seedling are organic matter, available potassium, moisture content, porosity, bulk density, N and P( P<0.05).

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Comparison between Betalain and Flavonol Accumulation in Tricolor Bougainvillea peruviana ‘Thimma’ Based on Transcriptome

XU Su-Xia, HUANG Qing-Yun, LIN Chun-Song, HUANG Yi-Jin, HU Ou

Bulletin of Botanical Research 2017, 37 (2): 249-258. DOI: 10.7525/j.issn.1673-5102.2017.02.013

Abstract （773）

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Bougainvillea peruviana ‘Thimma’, accumulating betalains, not anthocyanins occurring in majority of plants. The cultivar was characterized with either white, magenta, and white-magenta(variegated) bracts in the same branches. We first used magenta(Yp), bicolor(Ym) and white(Yw) open inflorescences for transcriptome analysis by sequencing. Candidate genes involving in pigment metablisms were validated by real-time PCR. We obtained 73,325 genes with average length of 616 bp. Out of DEGs, 327 candidate genes of betalain-biosynthesis, 308 flavonoid-biosynthesis genes and 466 anthocyanin-accumulation genes were detected. Eight candidate pigment-related genes were verified, four were responsible for betalain production(PPO, CYP76AD1, cDOPA-5-GT, DODA), and four responsible for flavonoids production(FLS, DFR, LDOX, 3-GT). Expression strength of four betalain production genes was well consistent with betalain-accumulation in Yp, Ym and Yw. Full later three anthocyanidin-producing genes(DFR, LDOX, 3-GT) were first verified and the expression varied in three samples. Expression of all eight candidate genes was well consistent with RNA-sequencing data. B.peruviana could provide a good tool to study physiological, phytochemical and molecular mechanisms in Bougainvillea, effectively eliminating the interference of some factors.

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Genetic Diversity and Fingerprints with SCoT Markers in Actinidia

ZHANG An-Shi, ZHANG Zhong-Hai, QI Xiu-Juan, LIU Ying, LUO Yang

Bulletin of Botanical Research 2017, 37 (2): 259-265. DOI: 10.7525/j.issn.1673-5102.2017.02.014

Abstract （762）

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SCoT were used to analysis genetic diversity of the samples of 32 Actinidia varieties. The 11 primers were screened from 47 SCoT primers and 185 SCoT bands were obtained, including 180 polymorphic bands, with a polymorphism rate of 97.30%. The average Nei's gene diversity( H) and Shannon's information index(I) were 0.2384 and 0.3778. The cluster analysis conducted with UPGMA showed that 32 Actinidia varieties were divided into 5 groups at the genetic similarity coefficient of 0.78, and the constructed phylogenetic trees based on SCoT were consistent with the morphology results. The DNA fingerprints for 32 Actinidia varieties were established with 16 sites from 4 SCoT primers, and 32 Actinidia varieties could be identified by SCoT fringerprints.

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Variation and Selection of Growth Character on the Young Third Generation of Pinus massoniana

ZHANG Zhen, YU Qi-Xin, TENG Guo-Xin, JIN Guo-Qing, FENG Zhong-Ping, ZHOU Zhi-Chun

Bulletin of Botanical Research 2017, 37 (2): 266-273. DOI: 10.7525/j.issn.1673-5102.2017.02.015

Abstract （953）

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We analyzed the genetic parameters, breeding value, accumulation and distribution of 2-year raw biomass with a half diallel cross among 6×6 on three generation young growth period of forest on Laoshan farm in Zhejiang Province. There were significant genetic differences among the tree height, ground diameter, crown breadth and number of living branch in the young period of Pinus massoniana. There were also significant genetic differences in the period of 2-year biomass between different hybrid combinations. The ground biomass accounted for 87.17% of the total plant biomass in 2-year hybrid combination. The height growth was the largest in the young period of tree growth trait. The height growth was increased by 4.23 times from 5 to 2 years. According to method of comprehensive breeding value, the tree height( H) breeding value combined with conjointly diameter( D ₀), and 2 a raw stem biomass index( B _s) to choose higher survival rate of hybrid combinations. The higher survival rate combinations were 22×44, 33×22 and 40×44 based on 20% year old open pollination progenies. Based on the value of tree height and ground diameter of plant breeding, 16 number of plant were chosen accounting for 2%. The average gain were 1.07 m(tree height), 1.58 cm(ground diameter), 0.32 m(crown breadth) and 4.67(living branch number), respectively.

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Growth Traits Variation Analysis and Family Selection of 3-year-old Various Ploidy Betula platyphylla

HUANG Hai-Jiao, PENG Ru-Sheng, LIU Yu, JIANG Jing

Bulletin of Botanical Research 2017, 37 (2): 274-280. DOI: 10.7525/j.issn.1673-5102.2017.02.016

Abstract （810）

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The growth traits analysis and family selection of tree progeny are important links in the tree breeding. To select excellent birch hybrid progeny, 21 Betula platyphylla families were employed to determine the height(H), diameter at breast height(DBH) and volume(V) of 3-year-old seedlings. Results of variance analysis showed that differences of all growth traits among different families were highly significant( P<0.01). Family heritability of H, DBH and V were all higher than 0.75, belonging to highly inheritance. The family 311-15 was excellent at H, DBH and V, and 9.09%, 11.25%, 40.46% higher than other families, respectively. Moreover, there were not significant differences between triploid families 311-9, 311-25, diploid families 211-7, 211-2, 211-3, 211-6, 211-1 and triploid family 311-15 at H, DBH and V. Based on the analysis of subordinate function, triploid families 311-15, 311-9 and diploid families 211-7, 211-2 were selected as excellent clones when the selection rate was 20.00%. The genetic gains of four clones on H, DBH and V were 5.24%, 7.18% and 19.72%, respectively. Taking the average volume+2S as standard, 52 superior individuals were screened out. These results provide a reference for the popularization of birch excellent family and superior individual selection at southwest of liaoning, also form an important basis for alterations of high generation triploid seed orchards.

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PSⅡ Function and Intrinsic Characteristics of Light-harvesting Pigment Molecules for Sun- and Shading-leaf in Magnolia grandiflora During Overwintering

HU Wen-Hai, YE Zi-Piao, YAN Xiao-Hong, YANG Xu-Sheng

Bulletin of Botanical Research 2017, 37 (2): 281-287. DOI: 10.7525/j.issn.1673-5102.2017.02.017

Abstract （745）

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Light absorption and energy transfer are determined by intrinsic characteristics of light-harvesting pigment molecules, which also have impacts on distribution of excited energy for photochemical reaction, heat dissipation and chlorophyll fluorescence. We compared the differences of the PSⅡ function and intrinsic characteristics of light-harvesting pigment molecules of sun- and shading-leaves to study the photoprotective strategies in overwintering Magnolia grandiflora. The slight photoinhibition was caused in leaves of M.grandiflora by low temperature during overwintering. Natural sunlight enhanced photoinhibition in sunleaf, however, low light condition was propitious to the recovery of photoinhibition in shading-leaf. Sun-leaf had lower chlorophyll content and the numbers of light-harvesting pigment molecules( N ₀) to reduce light energy absorption. Sun-leaf also possessed higher photochemical function and thermal energy dissipation in PSⅡ, which would protect photosynthetic apparatus against damage by low temperature and high light. Shading-leaf exhibited lower capability of photochemical reaction, however, possessed greater thermal energy dissipation, which would alleviate photoinhibition of shadingleaf under temporal high light condition during overwintering.

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Phenotypic Diversity Analysis of Seedling Leaf Traits of Manglietia conifera Dandy

WEN Shan-Na, ZHONG Chong-Lu, JIANG Qing-Bin, CHEN Yu, ZHANG Yong, LI Qing-Ying

Bulletin of Botanical Research 2017, 37 (2): 288-297. DOI: 10.7525/j.issn.1673-5102.2017.02.018

Abstract （4298）

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It studied the phenotypic variation in the leaves of Manglietia conifera with 12 provenances seedlings from Vietnam, and measured the leaf morphological traits(length, width, area, perimeter, length-width ratio, petiole length) and micro morphological characteristics of stomata(density, length, width, area, length-width ratio) by the variance analysis, variable coefficient(CV), correlation analysis and principal components analysis(PCA). There were significant differences in leaf phenotypic traits among M.conifera provenances. The average CV of leaf morphological traits of provenance LC2 was the maximum(22.09%), and TQ2 was the minimum(12.76%), leaf area recorded the largest CV(28.60%) while stomata width recorded the smallest CV(5.19%). The result of correlation analysis showed that there was significant correlation between leaf phenotypic traits( P<0.05), leaf perimeter was significantly correlated with longitude, and leaf length-width ratio were significantly correlated with longitude and latitude( P<0.05), leaf phenotypic traits were not significantly correlated with longitude and altitude)( P<0.05). By PCA, the cumulative contribution rate of the first 3 principal components was 89.29%, basically represent all the information of the original character. By cluster analysis, the 12 provenances could be clustered into 4 groups at the Euclidean distance of 5. There is abundant variation of leaf phenotypic traits in M.conifera provenances, longitude and latitude have obvious influence on leaf morphological characteristics of M.conifera, and there is difference of variation degree among provenances and leaf phenotypic traits of M.conifera. Our study provides theoretical basis for genetic improvement of M.conifera.

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Influences of Fertilization on Growth and Fruiting of Fraxinus mandschurica

WANG Xi-Gang, ZHAN Ya-Guang, ZHANG Gui-Qin, YAN Chao-Fu, LI Xiang, ZENG Fan-Suo

Bulletin of Botanical Research 2017, 37 (2): 298-303. DOI: 10.7525/j.issn.1673-5102.2017.02.019

Abstract （1174）

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The experiment was conducted to study the effect of fertilization on growth and flowering of Fraxinus mandschurica. Fertilizers with the orthogonal test containing 3 factors and 3 levels of N, P and K were formulated. The study materials included 2-year-old grafting seedlings of F.mandschurica, 13-year-old juvenile F.mandschurica and 29-year-old adult F.mandschurica. Height and ground diameter of grafting seedlings, height and DBH(diameter at breast height) of juvenile, flowering rate and seed setting rate of adult were measured. The results showed that fertilization promoted the growth of F.mandschurica. For grafting seedlings, the order of effect of 3 fertilizers on height growth and ground diameter growth is N > P > K, and the optimal combination is with N of 0.006 kg, P of 0.012 kg and K of 0.008 kg. For the juvenile, the order of effect of 3 fertilizers on height growth and diameter growth is N > P > K, and the optimal combination is with N of 0.15 kg, P of 0.3 kg and K of 0.2 kg. For adult trees, the order of effect of 3 fertilizers on flowering rate and seed setting rate is N > K > P, and the optimal combination is with N of 0.3 kg, P of 0.6 kg, and K of 0.4 kg. Therefore, the fertilization could improve the growth of F.mandschurica.

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Allocation Patterns and Allometric Relationships between Above- and Below-ground Biomass of Three Life-form Herbaceous Plants in the Gurbantünggüt Desert

ZHAO Jing, WANG Xi-Yuan

Bulletin of Botanical Research 2017, 37 (2): 304-311. DOI: 10.7525/j.issn.1673-5102.2017.02.020

Abstract （916）

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We took biomass survey samples on herbaceous plants along the southern margin Gurbantünggüt desert to study the biomass allocation features of herbaceous plants of different life-forms in Gurbantünggüt Desert. Results showed that:(1) most of the total biomass of the sampled 34 species of herbaceous plants was less than 4 g, most of whose above-ground biomass(AGB) and below-ground biomass(BGB) was less than 2 and 0.5 g, respectively, and their root-shoot ratios(R/S) were less than 0.25; (2) Ephemeroids allocated more biomass to below ground parts and had a higher R/S, which distributed around 1.67 intensively; Ephemerals and annuals allocated much less biomass to belowground parts, both of which has smaller R/S around 0.15; (3)The growth speed of the biomass of up-ground and below-ground parts of ephemeral parts was equal, but that of the ephemeroids was unequal. The proportion of the ephemeroids biomass allocated to the below-ground parts was increased complying with the individual's enlargement, while annuals biomass was on contrast. Therefore, the biomass of up-ground and below-ground parts of ephemeral, ephemeroids and annuals has different allocation features, which is consistent with the special plant life story features.

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