Bulletin of Botanical Research

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Rediscovery of Ulota germana(Mont.) Mitt.(Orthotrichaceae) In New Zealand

WANG Qing-Hua, JIA Yu

Bulletin of Botanical Research 2018, 38 (1): 1-6. DOI: 10.7525/j.issn.1673-5102.2018.01.001

Abstract （2774）

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Ulota germana(Mont.) Mitt., formly as U.rufula(Mitt.) A.Jaeger, was recorded from only Marlborough District of South Island in New Zealand and it has not been refound for more than 60 years. A new locality is added here: Nelson, Denniston, near parking area at top of Denniston Incline. The plants from New Zealand agree well with those from South America except some variation in the size of leaves, perichaetial leaves and spores. This new specimen is well described, especially the opercula and calyptra not seen in previous works, the distribution is mapped and its distinction from other close species is discussed here. Ulota luteola(Hook.f. & Wilson) Wijk & Margad., formly as U.pygmaeothecia(Müll. Hal.) Kindb. is excluded from the moss flora of New Zealand.

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Study on The Seed Biological Characteristics of Chimonanthus praecox of Zhejiang Province

DU Hui-Cong, ZHANG Ying, TIAN Min, WANG Cai-Xia

Bulletin of Botanical Research 2018, 38 (1): 7-13. DOI: 10.7525/j.issn.1673-5102.2018.01.002

Abstract （907）

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To investigate the seed biological characteristics of wild Chimonanthus praecox of Zhejiang Province and improve germination rate,which provided the reference for conservation and utilization.The result showed that the 1000-seed weight was about 396.98 g, water content of the seed was about 15.66%, the viability was about 91%.The germination inhibitor of Ch.praecox mainly exist in kernel,the inhibitory methanol crude extracts was stronger than aqueous crude extracts. The exist of dense and hard seed coat and germination inhibitor make the highest seed germination rate is less than the seed lots viability and also influence the germination uniformity.Seed of soaking after pricked can improve the seed structure and in favor of germination.

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Response Analysis of Larix sibirica to Climate Warming at Different Elevations in the Eastern Tianshan Mountains

ZHANG Qing, YU Rui-De, ZHENG Hong-Wei, YANG Mei-Lin, GAN Miao

Bulletin of Botanical Research 2018, 38 (1): 14-25. DOI: 10.7525/j.issn.1673-5102.2018.01.003

Abstract （1309）

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Under the background of climate warming, we discussed the response relationship between the radial growth of Larix sibirica and the climatic factors at different elevation of the eastern Tianshan Mountains, and validated this relationship by using the redundant analysis method. Compared with the standard chronology, the residual chronology of different elevation has larger characteristic parameters, higher SNR and mean sensitivity, and more environmental information, which is better to represent the overall change of trees. At 2 160 m(L1), after the rapid warming of the chronology(after 1985), the correlation between the tree-ring width and the temperature was reducing significantly. At 2 430 m(L2), the chronology is less associated with precipitation and temperature in two periods. At 2 700 m(L3), the positive response of the chronology to precipitation and temperature from 1985-2013 was enhanced. The response of radial growth to monthly climatic factors is higher in November and December of the last year, and the correlation between temperature and tree-ring width index is higher in June and July of the current year. Climate warming reduces the radial growth of trees, which is more obvious in low elevation areas. At all elevation, the sensitivity of L.sibirica to climate warming is decreasing. The radial growth of the L.sibirica at high elevation is mainly affected by the temperature, while that at middle and low elevation area is mainly affected by precipitation and temperature.

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The Different Altitude Gradient Change Rules of the Main Shrub Community in Arid Valleys of the Bailongjiang River with Different Slope

WANG Fei, TU Cai-Yun, CAO Xiu-Wen, LIU Jin-Qian, YANG Yong-Hong, ZHANG Tao, QI Hao

Bulletin of Botanical Research 2018, 38 (1): 26-36. DOI: 10.7525/j.issn.1673-5102.2018.01.004

Abstract （1121）

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The change rules of different slope on the structure characteristics and species diversity of the Main shrub community in arid valleys of the Bailongjiang River were studied with Quadrat survey method. Aim to understand the vegetation characteristic and Species diversity changes from different elevation gradient of arid valleys in bailongjiang river, It will provides the theory basis for vegetation restoration in this region.Results show that: (1)species are different in the same slope of different altitude gradient, as well as in the different slope of same altitude gradient. with the increase of altitude,the species of different slope show that increased after decreased first, The main vegetation types are also different in the same slope of different altitude gradient; (2)With the increase of altitude gradient, α-diversity of shrub community and α-diversity of herb community in different slope all showed that a trend from rise to decline . Correlation analysis was carried out on the α-diversity index, contribution rate of species diversity for the species diversity is the largest,it is showed that the richness index(D ₁, D ₂)> ecological dominance index(SN)> interspecific opportunity index(H)> evenness index of community(R); (3)With the change of altitude gradient, β-diversity Whittaker index of the main shrub community have less change in different slope, the maximum appeared at an altitude of 1 250-1 650 m,Routledge and Cody β-diversity index between an altitude of 1 450-1 650 m reache maximum, but generally presents the waveform changes. β-diversity of herb community have larger change, At an altitude of 1 050-1 250 m, β-diversity of herb community in the sunny slope reaches maximum. At an altitude of 1 250-1 450 m,β-diversity index of herb community in the semi-shady slope and semi-sunny slope reaches maximum, β-diversity index of herb community in the semi-shady slope and semi-sunny slope were higher than sunny slope. Species diversity α and diversity β are different that in Different slope directions and different altitudinal gradients in dry valleys of Bai Long River and different slope directions have a certain correlation with the change of altitudinal gradient, species diversity α and diversity β indicating that elevation and slope directions are one of the main factors affecting biodiversity.

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Analysis of Geographical Distribution of Abies in China under Climate Change

LIU Ran, WANG Chun-Jing, HE Jian, ZHANG Zhi-Xiang

Bulletin of Botanical Research 2018, 38 (1): 37-46. DOI: 10.7525/j.issn.1673-5102.2018.01.005

Abstract （3998）

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China is the country which has the most abundant and the widest geographical distribution of Abies Mill. species in the world. It is also the key area for studying the causes and law of Abies distribution. We collected the data of geographical distribution of Abies in China in three ways: Chinese Virtual Herbarium, Global Biodiversity Information Facility and relevant literatures. Based on the current and future climate scenarios, the maximum entropy model(MaxEnt) was used to simulate the potential distribution of Abies. Also, we analysed of habitat suitability and evaluated the protective capability of various regions in China, using GIS spatial analysis functions. The results showed that: (1)Southwestern Sichuan, northern Yunnan, as well as southeastern Tibet are the hot spots for Abies in China; (2)The area of suitable habitat of Abies in China will significantly reduce under the future climate change scenario; (3)The suitable habitat has the tendency of moving northwards in the future; (4)Yunnan province has the highest protective capability under the current climate scenario. Under the future climate change scenario, the protective capability of the western regions in China will increase, while that of the central and eastern regions will decline. This study can provide a theoretical basis and reference value for the protection of Abies.

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Seasonal Changes of the Input of Root Exudates and Its Driving Characteristics of Rhizosphere Microbe in a Cercidiphyllum japonicum Sieb. Plantation

WANG Xiao-Ping, XIAO Xiao, TANG Tian-Wen, LI Yun-Xiang, XIAO Juan

Bulletin of Botanical Research 2018, 38 (1): 47-55. DOI: 10.7525/j.issn.1673-5102.2018.01.006

Abstract （835）

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The rhizosphere processes involved in exudate have important ecological effects, but there is very little seasonal changes in situ of plants root exudatuions and their mediated soil biogeochemical cycles, especially endangered rare plant. With 30-year Cercidiphyllum japonicum Sieb.. plantation which is national secondary endangered plants in Dagou in west Sichuan, we used the in situ improved collecting device to study C.japonicum Sieb. root exudates in April, July, September and December 2014, collect rhizosphere and non-rhizosphere soil, and synchronously analyze the changes of rhizosphere microbiological properties. The results showed that:(1)the root secretion of dissolve organic carbon(DOC), total nitrogen(TN) rate appeared remarkable seasonal changes, the highest in summer and lowest in winter. (2)Rhizosphere of microbial biomass carbon(MBC), microbial biomass nitrogen(MBN) was significantly higher non-rhizosphere soil that showed the positive rhizosphere effect. The rhizosphere effect also showed a similar pattern of seasonal dynamics with the input of the root exudates, summer(July) is the highest, while winter(December) is the lowest. (3)By correlation analysis, there was a positive linear correlation between the DOC and TN rates in root exudates, and the rhizosphere effect of MBC, MBN rhizosphere effect and soil enzyme activity implied that the root secretion input was an important factor that could drive the activity of the rhizosphere microorganisms, and reveal the seasonal response of the input of the tree roots to the process and function of the rhizosphere microbial process. Future research should be strengthened in the coupling effect and mechanism about the root secretion input and the process of soil biogeochemical cycle.

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Histocytology Observation on the Somatic Embryogenesis in Herbaceous Peony Callus

WEI Dong-Xia, ZHANG Teng, ZHENG Yan-Yi, YU Xiao-Nan

Bulletin of Botanical Research 2018, 38 (1): 56-63. DOI: 10.7525/j.issn.1673-5102.2018.01.007

Abstract （874）

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The stem segments, leaves and petioles of three herbaceous peony was used as explants for somatic embryogenesis. Paraffin method was used for morphological and histological observation of somatic embryogeneris. The main conclusions were as follows: callus induction rate from stems of ‘Going Bananas’ was 100%. Proliferation rate of the callus was over 4.0. The best induction and proliferation medium of inducting callus was WPM+IAA 1.0 mg·L ^-1+6-BA 1.0 mg·L ^-1+NAA 0.5 mg·L ^-1+TDZ 0.5 mg·L ^-1+CH 0.625 g·L ^-1. Embryogenic callus generated after transferred the callus to 1/2 MS(Ca ²⁺ doubled)+2,4-D 2.0 mg·L ^-1+ABA 0.5 mg·L ^-1 or ZT 1.0 mg·L ^-1 medium of a constantly dark cultivation of 90 d. Globular embryos and heart embryos were generated after transferred the embryogenic callus to 1/2MS(Ca ²⁺ doubled)+6-BA1.0 mg·L ^-1+NAA 0.2 mg·L ^-1 medium for a constantly illumination cultivation of 60 d. There are exogenous and endogenous origins of somatic system. Exogenous includes origins from individual surface cells and origins from multiple subcritical cells. The difference of three ways lies mainly in starting position and starting cells number, they will all generated proembryo later. The splitting of embryogenic cells was symmetrical and the presence of asymmetric splitting cells was not found.

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Effects of Trichoderma asperellum Combined Application on Growth and Photosynthesis Characteristics of Populus davidiana×P.alba var. pyramidlis

LIU Zhao-Ying, JIANG Chuan-Ying, ZHAI Tong-Tong, CHANG Yuan, YAO Zhi-Hong, LIU Zhi-Hua, ZHANG Rong-Shu

Bulletin of Botanical Research 2018, 38 (1): 64-74. DOI: 10.7525/j.issn.1673-5102.2018.01.008

Abstract （987）

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In order to prepare compound fungi fertilizer of Trichoderma asperellum, we studied the effects of T.asperellum combined application on the growth and photosynthetic characteristics of Populus davidiana×P.alba var. pyramidlis(Shanxin poplar) by applying the concentration(5×10 ³ cfu·cm ^-3 soil) with different combinations of T.asperellum conidia T1(Ta536+Ta4), T2(Ta536+Ta4+Ta492) and T3(Ta536+Ta4+Ta492+Ta650) to roots of the tissue-cultured Shanxin poplar seedlings under field conditions. Analysis of variance showed that the induction time and different treatments had significant effects on height, basal stem, chlorophyll content, chlorophyll a/b value, P _n, Cond, C _i and T _r( P<0.05): the seedling heights, basal stems and dry weights of the treatment groups were higher than those of control group(CK) in different degree, and the effect was T3> T2> T1> CK; the biomass of poplar seedlings of T3, T2 and T1 separately increased 17.99%、14.28% and 10.54% compared with CK at 60 d; at 15 d, chlorophyll contents separately increased 7.79%, 6.91% and 4.17% and chlorophyll a/b values separately increased 7.79%, 5.84% and 4.73% compared with CK. In addition, the net photosynthetic rate( P _n), intercellular CO ₂ concentration( C _i), stomatal conductance( Cond) and transpiration rate( T _r) of the treatment were higher than those of CK, and the maximum net photosynthetic rate, apparent quantum efficiency and light saturation point of the treatment groups were higher than those of CK( P<0.05). At the same time, the interaction between the inducing durations and the different treatments levels of T.asperellum also significantly influenced the above indexes(except for P _n and C _i)( P<0.05). In brief, combination of 4 strains had synergistic effects that can enhance the adaptability of these strains to environment, promote growth and improve photosynthesis characteristics, and the T3 group has the biggest influence and the fastest action on the poplar seedlings.

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Protective Effects of Alternative Respiratory Pathway on Photosystem Ⅱ in Phaseolus vulgaris under CuCl ₂ Stress

XIE Jia-Jia, LU Li-Na, SHI Dai-Long, WANG Qing-Wen, JIA Ling-Yun, FENG Han-Qing

Bulletin of Botanical Research 2018, 38 (1): 75-80. DOI: 10.7525/j.issn.1673-5102.2018.01.009

Abstract （613）

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With “Nongpu” No.12 bean seedling leaves( Phaseolus vulgaris L.), we used the AOX inhibitor salicylhydroxamic acid(1 mmol·L ^-1; SHAM) to inhibit AOX activity in vivo, and studied the protective effects of alternative respiratory pathway on the photosystem Ⅱ under the CuCl ₂ stress. The treatment with CuCl ₂ caused a significant decrease in the ratio of F _v/ F _m(the potential maximal photochemical efficiency of PSⅡ), F _v'/ F _m'(the maximum quantum efficiency of PSⅡ photochemistry at illumination), Y(Ⅱ)(effective photochemical quantum yield of PSⅡ photosynthetic), qP(photochemical quenching coefficient), and the chlorophyll content, but the levels of NPQ(non-photochemical quenching) and the capacity of the alternative respiratory pathway were significantly increased. Compared with the bean seedling under CuCl ₂ stress, the CuCl ₂-stressed bean seedling pretreated with inhibitor of alternative respiratory(salicylhydroxamic acid) had lower levels of F _v/ F _m, F _v'/ F _m', Y(Ⅱ), qP and chlorophyll content, and higher levels of NPQ. These observations suggest that under the CuCl ₂ stress, the alternative respiratory pathway can protect the photosystem Ⅱ of the bean leaves from CuCl ₂ stress by alleviating the decrease of the photochemical efficiency of PSⅡ, maintaining the proportion of open PSⅡ centres, reducing the heat dissipation of the antenna pigment and chlorophyll content disruption.

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Effects of Drought Stress on the Growth and Biomass of Phoebe zhennan'seedling in Different Substrates Net Container

YAO Xiao-Lan, ZHOU Lin, FENG Mao-Song, HAO Jian-Feng, RAO Yuan-Lin, YAO Jun-Yu, QI Jin-Qiu

Bulletin of Botanical Research 2018, 38 (1): 81-90. DOI: 10.7525/j.issn.1673-5102.2018.01.010

Abstract （908）

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Adaptability and survival strategy of Phoebe zhennan in different substrates under periodic drought stress were studied to provide a reference for substrate selection of seedling cultivation to combat drought stress. According to the volume ratio, the seedlings of P.zhennan were cultured on five different substrates: T1: cultivated soil=10, T2: forest humus=10, T3: cultivated soil：forest humus：peat soil：pearlite=4：3：2：1, T4: cultivated soil：forest humus：peat soil：pearlite=3：2：4：1, T5: cultivated soil：forest humus：peat soil：pearlite=2：1：6：1. The drought stress gradients were designed randomly to study the effects of drought stress on the growth of P.zhennan seedling. The results were: (1)The physical and chemical properties of different substrates composition were closely related to the seedling growth and the seedling exhibited better growth performance in the substrates than traditional seedlings, but only T3 and T4 had better growth under heavy stress. (2)Along with the increase of the drought stress, the seedling height, basal diameter, leaf area, leaf area index, above ground biomass and underground biomass were aggravated, but the degree of influence was different. (3)The membership function value of the seedling quality was T3 >T4=T1 >T2 >T5, and T3 had the best quality and a strong tolerance of seedling to adapt to drought stress, improving the effectiveness of afforestation significantly. The suitable ratio of light substrates could coordinate the water, fertilizer, gas and heat, being propitious to the growth of the seedling in adversity.

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Transcriptomic Analysis of the Halostachys caspica in Response to Short-term Salt Stress

ZHANG Li-Li, ZHANG Fu-Chun

Bulletin of Botanical Research 2018, 38 (1): 91-99. DOI: 10.7525/j.issn.1673-5102.2018.01.011

Abstract （876）

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As a halophyte with strong salt tolerance, Halostachys caspica widely distributes in desert saline-alkali land. In order to reveal the genomic changes of gene expression under salt stress, transcriptome sequencing of H.caspica assimilating branches with treatments of 300 and 500 mmol·L ^-1 NaCl for 3 h was performed. A total of 153 298 unigenes with an average length of 643 bp were obtained with clean reads assembled. The 47 subclasses and 118 KEGG pathways were enriched in the GO terms and KEGG pathways, respectively. Differentially expressed genes analysis showed that there were 4 432 and 2 580 unigenes in response to low salt (300 mmol·L ^-1) and high salt (500 mmol·L ^-1) in short-term stress, respectively. The 1 245 unigenes were the common differentially expressed genes in the two salt stresses. They were mainly enriched in cellular process, metabolic process and response to stimulus. The osmotic regulation and reactive oxygen species (ROS) scavenging genes were screened out, and most of them were up-regulated. Therefore, H.caspica could improve short-term salt stress adaptation by enhancing the osmotic adjustment and ROS scavenging.

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Transcriptome Analysis of Dioscorea bulbifera L. Microtubers Conserved in vitro at Low Temperature

HONG Sen-Rong, WU Hui, ZHONG Lu, XIONG Si-Min, LUO Xia, LIU Xing

Bulletin of Botanical Research 2018, 38 (1): 100-109. DOI: 10.7525/j.issn.1673-5102.2018.01.012

Abstract （1025）

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With Dioscorea bulbifera L. microtubers, we studied the conservation in vitro at 4℃ low temperature and its transcriptome analysis. After the original data were pretreated, the effective ratio of reads in the control group(Con25) and the treatment group(Con4) was 98.67% and 98.69%, respectively. The 219792 transcripts of 200 bp(177Mb) and 161066 Unigenes(99Mb) were obtained after the removal of the splicing sequence; 10 species of the most NR annotations in the Unigene was Aureobasidium pullulans EXF-150, Vitis vinifera, Theobroma cacao, Oryza sativa Japonica Group, Amborella trichopoda, Setaria italica, Aureobasidium pullulans var. namibiae CBS 147.97, Jatropha curcas, Aureobasidium melanogenum CBS 110374 and Pestalotiopsis fici W106-1, respectively. The 5 KOG of the annotated most unigenes were general function prediction only, signal transduction mechanisms, posttranslational modification, protein turnover, chaperones, translation, ribosomal structure and biogenesis and energy production and conversion. The 5 pathway of the KEGG annotated most unigenes were carbon metabolism, biosynthesis of amino acids, glycolysis/gluconeogenesis, starch and sucrose metabolism and pyruvate metabolism. The number of annotated unigenes was 26 006, the number of different annotated enzymes was 1 177, and the number of different pathway was mapped to 327. Gene function of samples in Biological Process classification mainly gathered at cellular process and metabolic process, in Cellular Component classification it gathered at cell and cell part, in Molecular Function classification mainly concentrated in binding and catalytic activity. In conservation in vitro at low temperature of D.bulbifera L. microtubers, a total of 164 145 differentially expressed genes was obtained, of which 63305 genes were up-regulated and 100 840 genes were down regulated. Extremely significant GO terms of the differentially expressed genes were vacuole inheritance, single strand break repair, mitotic spindle elongation, maltose catabolic process, mannosyltransferase activity, mannosylphosphate transferase activity, cell wall mannoprotein biosynthetic process, G1 phase of mitotic cell cycle, incipient cellular bud site and establishment of mitotic spindle orientation. Extremely significant pathways of the differentially expressed genes were primary bile acid biosynthesis, steroid hormone biosynthesis, chlorocyclohexane and chlorobenzene degradation, bisphenol degradation, fluorobenzoate degradation, glycosaminoglycan biosynthesis-chondroitin sulfate, glycosaminoglycan biosynthesis-heparan sulfate, toluene degradation, naphthalene degradation and riboflavin metabolism. The experimental results provides a theoretical basis for germplasm conservation of D.bulbifera L. microtuber and its subsequent germination.

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Changes of Several Cations in Associated Plant Salicornia europaea and Suaeda stellatiflora in Different Growth Stages

ZHU Chuan-Ying, WU Wen-Bo, JIANG Zhen-Bo, WANG Yan-Qin

Bulletin of Botanical Research 2018, 38 (1): 110-116. DOI: 10.7525/j.issn.1673-5102.2018.01.013

Abstract （896）

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In order to analyze the change of Na ⁺, K ⁺, Ca ²⁺ and Mg ²⁺ in root, stem and leave at different growth stages of associated plants Salicornia europaea and Suaeda stellatiflora, these cations were determined by flame spectrophotometry and atomic absorption spectrometry. The result showed that the trends of Na ⁺, K ⁺, Ca ²⁺ and Mg ²⁺ were different in root and steam from leaf organs. The contents of Na ⁺ and K ⁺ were decreased in the root and stem, increased in leaf tissues with the development of plants. The accumulation of Ca ²⁺ and Mg ²⁺ were from low to high to reduce in roots and stem with two species growth and development, as same as in leaves of S.europaea. But accumulation of Ca ²⁺ and Mg ²⁺ were from high to low to up trend in leaves of S.stellatiflora with its growth and development. These two halophyte could absorb the Na ⁺ about 3% of roots, 5% of stems, and 10% of leaves of dry weight. The associated plants S.europaea and S.stellatiflora could become functional plants in improving saline-alkali soil, because these two plants have excellent potential to remove plenty of salt.

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Characteristic of Molecular Evolution for Lilium pumilum Redouté on Southeast of the Qinghai-Tibetan Plateau using nrDNA ITS and cpDNA petB/petD Sequence Analysis

JIANG Fu-Juan, TANG Nan, TANG Dao-Cheng, JU Xiu-Ting, SUN Jing-Qing

Bulletin of Botanical Research 2018, 38 (1): 117-124. DOI: 10.7525/j.issn.1673-5102.2018.01.014

Abstract （1006）

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In this study, total DNA was extracted from Lilium pumilum on southeast of the Qinghai-Tibetan Plateau using modified CTAB method. Using genome DNA as template, nrDNA ITS and cpDNA petB/petD regions were amplified, purified and sequenced. By sequence alignment, it was known that length of nrDNA ITS sequence was 696 bp, of which 4 variable sites with a percentage of 0.72% were found., The G+C content of the ITS sequence was 60.4% and 7 haplotypes were produced. The length of cpDNA petB/petD sequence was 616 bp, of which 1 was variable sites with a percentage of 0.16% was found. The G+C content of the sequence was 34.6% and 2 haplotypes were found. The petB/petD region of L.pumilum was more conserved than ITS sequences and evolved more slowly. The present distribution range of L.pumilum has probably experienced range expansion by the haplotype analysis of mismatch distribution and neutral test, and AMOVA analysis showed that the genetic variation of L.pumilum mainly existed within populations. The N _ST > G _ST( P>0.01)further showed that there was no significant phylogeographic structure of genetic variation in L.pumilum. Therefore, the nrDNA ITS sequence of L.pumilum was more suitable for phylogeographic study of this species.

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Expression and Function Analysis of Walnut JrGRAS2 Gene under Heat Stress

SU Li-Yuan, LI Xiao-Zhe, CHEN Shu-Wen, ZHANG Fang-Fang, ZHAO Ting-Ting, YANG Gui-Yan

Bulletin of Botanical Research 2018, 38 (1): 125-131. DOI: 10.7525/j.issn.1673-5102.2018.01.015

Abstract （935）

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The GRAS transcription factor is important for plant response to abiotic stress. To well understand the adaptive mechanism to adverse environment of walnut tree, a GRAS gene was cloned from the transcriptome of Juglans regia(Named as JrGRAS2). The expression of JrGRAS2 was analyzed under heat stress, and JrGRAS2 was inserted into the yeast expression vector pYES2 for constructing recombinant plasmid pYES2-JrGRAS2, which was transformed into a Saccharomyces cerevisiae strain(INVSCI). The yeast transformed with empty pYES2 was used as a negative control. The yeast expression system was used for analysis the heat stress tolerance. The full length open reading frame(ORF) of JrGRAS2 was 1 296 bp, the deduced protein was 47 405.83 Da with 431 amino acids, and the theoretical isoelectric point(pI) was 5.66. Under heat stress, JrGRAS2 was highly induced, especially exposed to 36℃ for 0.5 h in the stems, it was induced to 335.5-fold of control. When both recombinant yeasts were treated with 53℃, the JrGRAS2 expressed yeast displayed higher vitality and survival rate than the control yeast. Therefore, JrGRAS2 gene could effectively response to heat stress and improve heat tolerance of transgenic yeasts, JrGRAS2 may be an important candidate gene for walnut response to adverse stimulus.

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Cloning and Expression Analysis of S-Adenosine Methionine Decarboxylase( ThSAMDC) Gene from Tamarix ramosissima

ZHANG Yu, ZHANG Yue, ZHANG Chun-Rui, WANG Yan-Min, WANG Yu-Cheng, WANG Chao

Bulletin of Botanical Research 2018, 38 (1): 132-140. DOI: 10.7525/j.issn.1673-5102.2018.01.016

Abstract （887）

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S-adenosyl-L-methionine decarboxylase( SAMDC) is a rate-limiting enzyme that regulates the physiological and biochemical processes of plants by participating in the metabolic pathway of polyamines. The cDNA of S-Adenosine Methionine Decarboxylase gene(named ThSAMDC) was isolated and cloned by analyzing the transcriptome data of Tamarix hispida. The full-length of ThSAMDC is 2 085 bp, with three open reading frames, tiny ORF(tORF), upstream ORF(uORF) and main ORF(mORF). The mORF was 1 107 bp encoding 369 amino acids. The relative molecular weight and isoelectric points(PI) of the putative protein were 40.34 kD and 4.72. ThSAMDC-encoded protein has a number of strong hydrophilic regions, no obvious transmembrane region. Compared to the amino acid multiple sequence alignment of several other species, ThSAMDC includes two highly conserved domains: the proenzyme cleavage site(LSESSLF) and the PEST domain(TIHVTPEDGFSYAS) associated with the rapid degradation of the protein. Phylogenetic tree analysis shows that ThSAMDC has higher sequence similarity of 77% identities to the Spinach(SoSAMDC). Quantitative real-time PCR assay revealed that the mRNA level of ThSAMDC was significantly up-regulated under NaCl, PEG, ABA and CdCl ₂ treatments in T.hispida, suggesting that ThSAMDC may play an important role in drought resistance, salt tolerance and other stresses.

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SNP Sites Developed by Specific Length Amplification Fragment Sequencing(SLAF-seq) and Genetic Analysis in Ammopitanthus mongolicus

DUAN Yi-Zhong, WANG Jian-Wu, DU Zhong-Yu, KANG Fu-Ren

Bulletin of Botanical Research 2018, 38 (1): 141-147. DOI: 10.7525/j.issn.1673-5102.2018.01.017

Abstract （944）

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Ten site of Ammopiptanthus mongolicus distributed in Inner Mongolia, Ningxia and Gansu Province were used as experiment materials for sequencing by specific-locus amplified fragment sequencing(SLAF-seq). The scheme of the experiment was designed based on bio-informatics technology. Taking Glycine max as the reference genome, specific size of DNA were chosen to construct the SLAF-seq library. After high-throughput sequencing, a great amount of sequences were obtained and used to obtain the polymorphism SLAF tags by software alignment, then found the distribution of specific SNP sites. In total, we obtained 374 265 SLAF tags, including 56 295 polymorphic SLAF tags. According to these conditions that integrity >0.5 and MAF>0.05 screening from all SNP, obtained 102 025 group SNP which have high consistency. Phylogenetic relationship population structure and PCA analysis of ten individuals of A.mongolicus were analyzed using mathematical statistic method based on screening SNP, from the level of the genome to reveal the genetic differentiation between different groups. The results show that these A.mongolicus in ten different sites all originate from the same ancestor, but due to these factor that geographical location make the A.mongolicus produced genetic differentiation. Therefore, A.mongolicus growing in Inner Mongolia have close relationship, and that distributed in Ningxia and Gansu Province have close relationship.

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Arabidopsis thaliana Transcription Factor Ethylene-insensitive3(EIN3) Inhibits the Synthesis of Anthocyanins

XU Meng-Ke, LI Dan, MENG Lai-Sheng, JIANG Ji-Hong

Bulletin of Botanical Research 2018, 38 (1): 148-154. DOI: 10.7525/j.issn.1673-5102.2018.01.018

Abstract （1721）

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Ethylene-insensitive3(EIN3) and EIN3-like1(EIL1) proteins are important nuclear transcription factors in ethylene signal transduction pathways in higher plants. In plants, anthocyanins are one of the water-soluble natural pigments, and they play an important role in the growth and development of plants. We used the Arabidopsis thaliana double mutant ein3-1 eil1-3 as the research material. Firstly, we identified the mutants of ein3-1 eil1-3 and ein3-1 by RT-PCR. The direct observation indicated that the seeds and leaves of the ein3-1 eil1-3 were purple. By ultraviolet spectrophotometer analysis, the anthocyanin accumulation in ein3-1 eil1-3 mutant was significantly higher than that in the ein3-1 mutant and wild-type. GUS staining showed that EIN3 was mainly expressed in flowers, stigma, mature pollen, seed embryo and siliques, which was consistent with that anthocyanin accumulation of seeds and leaves was high in mutant ein3-1 eil1-3. Thus, A.thaliana transcription factor EIN3 as well as EIL1 is involved in the suppression of the anthocyanin synthesis.

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Study on Orthogonal Optimization and Antioxidation of Polysaccharide from Tara Seeds

YUAN De-Cheng, ZHAO Han-Mei, YANG Feng-Jian

Bulletin of Botanical Research 2018, 38 (1): 155-160. DOI: 10.7525/j.issn.1673-5102.2018.01.019

Abstract （796）

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Tara seed as raw material, process and antioxidant properties of protein and polysaccharide of Tara seed polysaccharide of Tara.Using the loss rate of polysaccharide and protein removal rate as the evaluation index to compare Sevage method, three trichloroacetic acid method and papain method of Tara polysaccharide deproteinization effect. Removing protein were optimized by using orthogonal optimization experimental design principle and the method of orthogonal analysis of four factors and three levels of papain method. The results showed that: Tara polysaccharide best protein removal technology for enzyme dosage 0.15 mL, hydrolysis time 90 min, enzymolysis temperature of 60℃, enzymolysis pH=6, protein removal rate was 95.19%, rate of polysaccharide is 75.02%. Through the study of Tara polysaccharide antioxidation of polysaccharide retention at Tara, it is showed thatthe total antioxidant activity of sugar was better, which had a strong scavenging effect on DPPH free radical.

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Lectotypifications of Names in Sympetalae from China-Ⅳ.

BI Hai-Yan, SHAO Qing, SUN Qian, YANG Zhi-Rong, LIN Yun, YUN Ying-Xia

Bulletin of Botanical Research 2018, 38 (2): 161-167. DOI: 10.7525/j.issn.1673-5102.2018.02.001

Abstract （4324）

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In type specimens deposited at Herbarium (PE), Institute of Botany, Chinese Academy of Sciences, the types of some taxon names in Sympetalae from China are found to be syntypes under Article 9.5 of International Code of Nomenclature for algae, fungi, and plants (Melbourne Code), including Lonicera chrysantha Turcz. var. longipes Maxim., L.syringantha Maxim., L.tangutica Maxim. and Viburnum erubescens Wall. var. carnosulum W.W. Smith in Caprifoliaceae, Valeriana minutiflora Hand.-Mazz. and V.tangutica Batal. in Valerianaceae, Scabiosa bretschneideri Batal. in Dipsacaceae, and Ainsliaea scabrida Dunn, Anaphalis hancockii Maxim., A.lactea Maxim., Artemisia smithii Mattf., Blumea henryi Dunn, Cacalia matsudae Kitam., Cremanthodium discoideum Maxim., C.humile Maxim., Leontopodium haplophylloides Hand.-Mazz., L.roseum Hand.-Mazz., Nannoglottis carpesioides Maxim., Saussurea quercifolia W.W.Smith var. major Anth., Senecio begoniaefolius Franch., S.koualapensis Franch., S.sagitta Maxim., S.tanguticus Maxim. and Tanacetum potaninii Krash. var. suffruticosum Krash. in Asteraceae. According to Article 8.1, 9.11 and 9.12, and Recommendation 9A.3, lectotypes for twenty-four names are here designated.

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The taxonomic study of Haplodontium Hampe(Bryaceae,Bryophyta) in China

WANG Xiao-Rui, LI Min, NIU Yu-Lu, ZHAO Jian-Cheng

Bulletin of Botanical Research 2018, 38 (2): 168-181. DOI: 10.7525/j.issn.1673-5102.2018.02.002

Abstract （710）

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Haplodontium Hampe, a new genus to China is described. Currently, there are three species of Haplodontium in China. Haplodontium zangii X.R.Wang & J.C.Zhao is a new species from Xizang, China. Haplodontium himalayanum(Mitt.) X.R.Wang & J.C.Zhao and H. sinensis(Dix.) X.R.Wang & J.C.Zhao are transferred to the genus Haplodontium from Mielichhoferia. The illustration and micrograph under light microscope(LM) and scanning electron microscope(SEM) of each species of Haplodontium in China are provided. Main morphological characters, habitat and geographical distribution, and morphological comparison with related species of them are discussed.

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Sporogenesis and Gametogenesis of Lagerstroemia speciosa

ZHU Li-Qiong, YUAN Juan, WEI Li-Di, WEI Yan-Chun, ZHAO Li-Jun

Bulletin of Botanical Research 2018, 38 (2): 182-188. DOI: 10.7525/j.issn.1673-5102.2018.02.003

Abstract （1107）

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The flower anatomical structures of Lagerstroemia speciosa in different development stages were studied by using method of paraffin section. The formation of the sporophyte and the development process of male and female gametophytes were also surveyed. The results showed that anther has four chambers. The anther wall shows a dicotyledonous type, which be comprised of epidermis, endothecium, middle layer and glandular tapetum. Cytokinesis occurs simultaneously with meiosis in a microspore mother cell, and the formed tetrads are mostly tetrahedral while some are decussate type. The mature pollen grains are of a 3-cell type with three grooves, occasionally aborted. The gynoecium was composed of 6 7 carpels. The ovary has 6 7 locules and there are several anatropous ovules in each locule. The ovules are crassinucellate and bitegmic. The megaspore mother cell divides to a linear megaspore tetrad. The chalazal megaspore develops into a polygonum type embryo sac. The mature embryo sac contains seven cells and eight cell nucleus. The most pollen and embryo sac development are normal, and L.speciosa can be used as a fine female parent in the hybridization breeding. According to the flower morphological characteristics in different blooming phenophase, the process of megaspore and microspore formation and development of male and female gametophytes can be determined initially.

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Taxonomic Position of Viola tuberifera Based on Morphological Variation of the Populations and Digital Specimen

SU Xue, HOU Yun-Yun, GAO Ting, WANG Ya-Li, SUN Kun

Bulletin of Botanical Research 2018, 38 (2): 189-194. DOI: 10.7525/j.issn.1673-5102.2018.02.004

Abstract （846）

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Viola tuberifera, which is endemic to the Qinghai-Tibetan Plateau and its adjacent region, is a typical dimorphic cleistogamous plant, and the taxonomical position of V.tuberifera is still controversial. We analyzed the morphological variation of leaves from the digital specimen and three sympatric natural populations of V.bulbosa and V.tuberifera. The PCA analysis of digital specimen indicated that all the samples were divided into two clusters, represented for V.tuberifera and V.bulbosa, respectively. The PCA analysis of sympatric natural populations showed the same results. The parameters reflected the characters of leaf apex and base, such as ΔW, AL, BL and LWR, are relatively lower variation within and among populations of V.tuberifera and V.bulbosa, but the variation of these parameters between V.bulbosa and V.tuberifera is significant( P<0.01). V.tuberifera and V.bulbosa should be treated as different taxon, and indicating the leaf morphological characters can be used in the classificatiopn of V.tuberifera and V.bulbosa.

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Species Diversity of Carpinus cordata Communities in Lishan Nature Reserve,Shanxi Province

WU Ping-Ping

Bulletin of Botanical Research 2018, 38 (2): 195-200. DOI: 10.7525/j.issn.1673-5102.2018.02.005

Abstract （732）

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With the dataset in the investigated field, we studied the species diversity of Carpinus cordata communities in Lishan Nature Reserve, Shanxi Province by using Patric index( R ₀), Simpson index( λ), Shannon-Wiener index( H'), Pielou index( E ₁) and Alatalo index( E ₅). The change trends of R ₀, H', E ₁ and E ₂ was almost similarity, and the change trends of λ with H' was positive. Overall, the rank of species diversity indices of C.cordata communities was Ass. Ⅰ Carpinus cordata-Lindera glauca-Phlomis umbrosa > Ass. Ⅲ Carpinus cordata-Forsythia suspense-Phlomis umbrosa > Ass. Ⅳ Carpinus cordata-Spiraea pubescens+Forsythia suspense-Smilacina japonica > Ass. Ⅶ Carpinus cordata-Sambucus williamsii-Thalictrum baicalense+Epimedium brevicornu > Ass. Ⅱ Carpinus cordata-Deutzia parviflora-Phlomis umbrosa > Ass. Ⅵ Carpinus cordata-Deutzia parviflora-Osmorhiza aristata > Ass. Ⅴ Carpinus cordata-Deutzia parviflora-Smilacina japonica. The diversity indices between the tree layer and shrub layer was negative correlation( P<0.05),and the diversity indices between the tree layer and herb layer was negative correlation( P<0.05) also, moreover, the diversity indices between the tree layer and herb layer was negative correlation( P>0.05). The differences among those associations of the C.cordata communities was not significant( P>0.05).

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Effects of Growth Environment and Tree Age on Biomass Allocation within Fruit Twigs of Fengdan( Paeonia ostii)

MA Han-Ze, WANG Cheng-Zhong, LI Ding, FAN Guo-Zhen, HU Yong-Hong, SONG Zhi-Ping

Bulletin of Botanical Research 2018, 38 (2): 201-211. DOI: 10.7525/j.issn.1673-5102.2018.02.006

Abstract （851）

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Biomass allocation is critical for understanding the yield formation of crops. Fengdan is the cultivated type originated from Paeonia ostii and the predominant variety of oil tree Paeonia that is newly cultivated as one of the oil tree crops in China. In order to examine the effects of growth environment and tree age on twig vegetative and reproductive growth, we compared the biomass characteristics of fruit twigs among eight 6-year-old Fengdan populations from different sites and that between 4, 6 and 8-year populations cultivated at Shanghai, and Tongling of Anhui Province. The number of fruit twigs increased with age increase, while there was no significant difference of twig growth and reproduction between different aged populations, indicating age effects existed at individual level but not twig level. There were significant correlations between the trait measurements, whereas, such correlations varied among populations. The body sizes and biomass allocations of fruit twigs were different among different populations, and the variations of biomass allocation associated with environmental variables. By allometric relationship modeling analysis, the fruit twigs from most of populations displayed constant biomass allocation between growth and reproduction functions(slope=1) with varied intercepts among populations. Therefore, the cultivation environment has significant effects on the growth and reproduction of Fengdan.

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Mega-and Microsporogenesis and Development of Female and Male Gametophytes in Michelia maudiae Dunn

XIONG Hai-Yan, LIU Zhi-Xiong

Bulletin of Botanical Research 2018, 38 (2): 212-217. DOI: 10.7525/j.issn.1673-5102.2018.02.007

Abstract （958）

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The mega-and microsporogenesis and development of female and male gametophytes in Michelia maudiae Dunn were investigated by using the paraffin section method. Anthers are tetradymous and the anther wall is composed of 5-7 layer of cells, Glandular tapetum, Cytokinesis in meiosis is modified simultaneous type, the tetrads are tetrahedral, isobilateral and decusate. Mature pollen grains are 2-cell type. The ovule of M.maudiae is anatropous bitegmic and nucellus thickly. The megaspore mother cells undergo meiosis divides into linear type tetrads, and the one in the chalazal end is functional, Polygonum type. The female and male gametophytes in spring-flowering M.maudiae were developed normally, heterogamy. The megaspore mothercells formed after the petals and stamen were withered. The development of microspore in autumn is in the period of sporozoic cell or primary spore, abortionthen. Megaspore in autumn-flowering M.maudiae undeveloped when flowering, withers away after flowering, were mostly developed unnormally. The study had accumulated information on the reproduction and hybridization in M.maudiae Dunn.

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Effects of Phosphorus Supply on Growth and Foliar Nutrition of Dalbergia odorifera Seedlings

ZHAO Xia, XU Da-Ping, LIU Xiao-Jin, ZHANG Ning-Nan, YANG Zeng-Jiang

Bulletin of Botanical Research 2018, 38 (2): 218-224. DOI: 10.7525/j.issn.1673-5102.2018.02.008

Abstract （800）

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A pot trial was conducted for Dalbergia odorifera seedlings with fertilization at levels of 0, 15, 30, 60, 100 and 150 mg P per seedling in a completely randomized block design, and seedling growth performance, biomass and foliar nutrient contents were measured and analyzed. Suitable phosphorus application was finally determined for growing seedlings of this species by critical level approach. Growth and foliar nutrient of D.odorifera seedlings were studied under different levels phosphorus addition to reveal the phosphorus demand and determine the optimal phosphorus supply for its seedlings. The results showed that addition of phosphorus fertilizer could enhance growth and biomass accumulation of D.odorifera seedlings. With the increase of phosphorus supply, seedling height, root collar diameter, lead area and biomass increased first and then declined, and their peaking values occurred at the fertilizer level of 60 mg P per seedling, which were 3.07, 3.35, 49.21 and 24.25 times of those of control. P fertilization decreased foliar nitrogen and potassium contents, while increased foliar phosphorus and magnesium contents, and foliar P contents showed no significant difference among fertilization treatments of 30, 60 and 100 mg P per seedling, which were about 1.65 times of that of control. Taking parabolic relationship between seedling biomass and foliar phosphorus content, content ratios of nitrogen to potassium and phosphorus to potassium, the optimal foliar phosphorus content ranged from 1.35-2.32 g·kg ^-1, and 60-100 mg P per seedling was determined as the optimal phosphorus supply for D.odorifera seedlings by critical level approach.

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Effects of Fertilization on the Vegetative and Reproductive Growth of Dalbergia odorifera T.Chen.

WANG Li-Yun, LIU Xiao-Jin, CUI Zhi-Yi, XU Da-Ping, YANG Zeng-Jiang, ZHANG Ning-Nan, HONG Zhou, CHEN Jia-Tang, HUANG Wei-Rong

Bulletin of Botanical Research 2018, 38 (2): 225-231. DOI: 10.7525/j.issn.1673-5102.2018.02.009

Abstract （1041）

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The experiment was conducted to study the fertilization on vegetative and reproductive growth of Dalbergia odorifera, and then provide useful suggestions for plantation management. The effects of Nitrogen(N, 185.6 g N per tree), Phosphorus(P, 120 g P ₂O ₅ per tree), Potassium(K, 120 g K ₂O per tree) as well as their mixtures(PK, 120 g P ₂O ₅ mixed with 120 g K ₂O per tree; NPK, 185.6 g N mixed with 120 g P ₂O ₅ and 120 g K ₂O per tree) on leaf growth, branch development, inflorescences set and biomass accumulation of a 8-year-old D.odorifera plantation were studied. There were significant differences on vegetative and reproductive growth of D.odorifera among these treatments( P<0.05). Nitrogen(N) treatment was helpful for vegetative growth of D.odorifera, and suitable for large-timber forest cultivation, because the ratio of vegetative branch, leaf number and leaflet width of vegetative branches were increased by 40.25%, 21.75% and 9.52%, and the inflorescence diameter and biomass were decreased by 12.75% and 48.63%, respectively. While P and P mixed with K were more appropriated for breeding purpose, because the ratio of vegetative branch was decreased by 47.96% and 46.84%, respectively. When N, P and K were mixed together(NPK), the vegetative and reproductive growth were both enhanced significantly( P<0.05), because the ratio of vegetative branch, length of vegetative branch, leaf number, leaflet length, leaflet width, branch biomass, inflorescence number and inflorescence biomass were increased by 26.04%, 68.16%, 32.98%, 15.20%, 11.40%, 83.60%, 54.20% and 49.84%, respectively. Thus, the vegetative and reproductive growth of D.odorifera can be regulated though formula fertilization(biased to N or P and K).

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A Preliminary Study on the Application of Self Pollination in Primula merrilliana Flower Color Selection

LIU Jin, XIA Qi-Ping, WANG Jian, LI Zhen, SHAO Jian-Wen

Bulletin of Botanical Research 2018, 38 (2): 232-237. DOI: 10.7525/j.issn.1673-5102.2018.02.010

Abstract （1001）

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Primula merrilliana is a rare ornamental flowers species and endemic to China. We preliminarily studied the application of self pollination in the selection and breeding of its flower color. The compatibility of self pollination in P.merrilliana was relatively higher, the average number of seeds was up to 36.03 per fruit, which reached 71.11% of the seeds of legitimate pollination(50.69 seed per fruit), and the germination rate of self pollination seeds was up to 41.13%, which was 57.98% of seeds germination rate of legitimate pollination. The analysis of flower color showed that the mean value of R of self pollination progeny population(0.382) was significantly higher than that of the parental population and the progeny population of legitimate pollination(both was about 0.375).Therefore, we should pay attention to the application of self pollination in the breeding of P.merrilliana because it is beneficial for flower color selection and obtaining of homozygous lines, although there is a certain level of inbreeding depression.

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GC/MS Metabonomics Analysis of Dioscorea bulbifera L. Microtubers Conserved in vitro at Low Temperature

YIN Ming-Hua, DENG Hong-Gen, JIANG Yan, WAN Lin, WU Li-Xia, LING Fei, WANG Jin-Hua

Bulletin of Botanical Research 2018, 38 (2): 238-246. DOI: 10.7525/j.issn.1673-5102.2018.02.011

Abstract （1165）

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We applied GC/MS detection method to different metabolites of Dioscorea bulbifera L. microtubers conserved in vitro at low temperature. Compared with D.bulbifera L. microtubers conserved in vitro at 25℃, different metabolites of D.bulbifera L. microtubers conserved in vitro at 4℃ had alanine, catechin, N,N-Di-(2-Hydroxyethyl)-methanamine, salicylic acid, citric acid and sorbose. In the conservation in vitro of microtuber at 4℃, alanine mainly involved in cyanoamino acid metabolism; Catechin mainly involved in biosynthesis of secondary metabolites, flavonoid biosynthesis and biosynthesis of phenylpropanoids; Salicylic acid mainly involved in polycyclic aromatic hydrocarbon degradation, microbial metabolism in diverse environments, plant hormone signal transduction, biosynthesis of secondary metabolites, dioxin degradation, phenylalanine metabolism, degradation of aromatic compounds, biosynthesis of plant hormones, biosynthesis of siderophore group non-ribosomal peptides and biosynthesis of phenylpropanoids; Citric acid mainly involved in biosynthesis of alkaloids derived from ornithine, lysine and nicotinic acid, biosynthesis of alkaloids derived from histidine and purine, microbial metabolism in diverse environments, biosynthesis of plant secondary metabolites, 2-Oxocarboxylic acid metabolism, biosynthesis of terpenoids and steroids, carbon fixation pathways in prokaryotes, biosynthesis of secondary metabolites, biosynthesis of alkaloids derived from shikimate pathway, biosynthesis of alkaloids derived from terpenoid and polyketide, citrate cycle(TCA cycle), biosynthesis of plant hormones, glyoxylate and dicarboxylate metabolism, two-component system, biosynthesis of phenylpropanoids and biosynthesis of alkaloids derived from ornithine, lysine and nicotinic acid. The preliminary findings of different metabolites of D.bulbifera L. microtubers conserved in vitro at low temperature would lay the foundation for further understanding the molecular mechanism of conservation in vitro at low temperature and provide a theoretical basis for low temperature conservation breaking dormancy and its subsequent germination D.bulbifera L. microtubers.

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Analysis of BpBEE1 Gene Promoter in Betula platyphylla

XU Si-Jia, YAN Bin, DONG Jing-Xiang, WU Dan-Yang, LI Hui-Yu

Bulletin of Botanical Research 2018, 38 (2): 247-253. DOI: 10.7525/j.issn.1673-5102.2018.02.012

Abstract （824）

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Brassinolide Enhanced Expression( BEE) genes belong to bHLH transcription factors family, and were involved in Brassinolide signal pathway. In this study, the promoter of BpBEE1 gene was cloned, and regulatory elements in the promoter regions were analyzed using the online program PLACE. The results showed that the BEE1 promoter sequence contained various hormone responses elements. The results showed that the GUS activity was increased in the veins and roots, and the activity of promoter was enhanced after MeJA, SA, BL and ABA treatment, especially MeJA, SA and BL treatment. We propose that BpBEE1 is involved in hormone response, and play a role in the process of plant growth and development.

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Arabidopsis thaliana Transcription Coactivator ANGUSTIFOLIA3(AN3) in Regulating the Stamen Formation

LI Dan, XU Meng-Ke, JIANG Ji-Hong, MENG Lai-Sheng

Bulletin of Botanical Research 2018, 38 (2): 254-259. DOI: 10.7525/j.issn.1673-5102.2018.02.013

Abstract （884）

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Stamens are important reproductive organs which produce pollen in seed plants, and their normal development is related to the reproductive status of plants and affects the yield of crops. Two knockout mutants an3-1 and an3-4 of Arabidopsis thaliana transcription coactivator ANGUSTIFOLIA3(AN3) were identified by RT-PCR. By morphological examination, the stamens of the an3-1 and an3-4 mutants were shorter than those of wild-type, whereas, the pistils were not changed significantly. By constructing AN3 promoter with GUS expression vector, the flower tissues of obtained Pro-AN3-GUS plants were stained and observed. The AN3 promoter in Arabidopsis seed embryos, mature pollen, stigma, petals was expressed. AN3 could be expressed in important tissues such as floral organs during Arabidopsis reproductive growth. These above results are consistent with the short stamens of an3-1 and an3-4 mutants. Thus, AN3 positively regulates the stamen development of A.thaliana.

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Construction and Functional Verification of Plant Expression Vector of Tamarix ThDUF106 Gene

GE Xiao-Lan, LIU Cai-Xia, ZHANG Xin-Xin, LI Ying, QU Guan-Zheng

Bulletin of Botanical Research 2018, 38 (2): 260-267. DOI: 10.7525/j.issn.1673-5102.2018.02.014

Abstract （903）

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ThDUF106 gene were involved in the stress tolerance of Tamarix hispida, and the expression of ThDUF106 was up-regulated in T.hispida under salt-alkali stress. In order to study the function of ThDUF106 gene in T.hispida, the temporal and spatial expression of these gene was detected by qRT-PCR in the leaves and roots of three-month T.hispida under abiotic stress such as salinity, drought and heavy metals. The gene encoding this gene was also analyzed, and the gene was constructed and transformed into tobacco by the plant expression vector. Transgenic tobacco lines and wild-type one-month seedlings were treated, and physiological and biochemical indexes were compared and analyzed. The antioxidant status of transgenic and wild type tobacco was basically the same under various duress, indicating that the gene was not expressed in tobacco and could not improve the antioxidation of tobacco.

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Cloning and Bioinformatics Analysis of Two bZIP Genes of Bambusa emeiensis and Their Induced Expression under Abiotic Stresses

GONG Dao-Yong, HU Shang-Lian, CAO Ying, LU Xue-Qin, ZHANG Qing-Bo

Bulletin of Botanical Research 2018, 38 (2): 268-277. DOI: 10.7525/j.issn.1673-5102.2018.02.015

Abstract （870）

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According to the transcriptome data of Bambusa emeiensis shoot, two bZIP genes designated as BebZIP2 and BebZIP6 were cloned and their bioinformatics were analyzed. The bioinformatics analysis results showed that the full-length cDNA sequence of BebZIP2 and BebZIP6 were 504 and 720 bp, and encoded 167 and 239 amino acids, respectively. BebZIPs and rice OsbZIP52/RISB5 proteins were clustered into the same branch, which were bZIP-related proteins. The tissue expression patterns of two BebZIP genes were analyzed by real-time PCR. Two BebZIP gene sequences were expressed in all shoots, stalks, unfolding leaves and rolled leaves of B.emeiensis. There were the expression differences of the same gene in the different tissues and expression quantity in the descending order was unfolding leaves, rolled leaves, stalks and shoots. The seedlings of B.emeiensis were treated with ABA, NaCl and PEG 6000 for abiotic stress analysis. The expression level of BebZIP2 and BebZIP6 genes treated by salt, drought and ABA stresses have different degree of sensitivity.

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Gnentic Diversity and Phylogeographic Structure of Triosteum pinnatifidum based Chloroplast DNA Sequence rbcL-accD

LIU Hai-Rui, GAO Qing-Bo, ZHANG Fa-Qi, XING Rui, CHI Xiao-Feng, CHEN Shi-Long

Bulletin of Botanical Research 2018, 38 (2): 278-283. DOI: 10.7525/j.issn.1673-5102.2018.02.016

Abstract （790）

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Triosteum pinnatifidum is perennial herb belonging to Triosteum (Caprifoliaceae) with the distribution in northwest Qinghai-Tibetan Plateau and adjacent mountain regions and Japan. It's endemic species of East Asia. We studied the sequence features, genetic variation and phylogeographic structure of T.pinnatifidum based on chloroplast DNA intergenic region rbcL-accD. Analysis of molecular variance for the whole populations revealed that 72.28% of total genetic variation occurred among populations, and 27.72% within populations. Dominant haplotype H2 had wide distribution while the private haplotypes could be found in different populations. There was no phylogeographic structure( N _ST< G _ST) and low gene flow( N _m=0.096), indicating the geographic isolation and variable habitat created by the special topography or ecological environment on the Qinghai-Tibetan Plateau and adjacent mountain regions may form current distribution of haplotypes. T.pinnatifidum experienced nonsignificant species selection or expansion. Indeed, we speculate there were different refugias for T.pinnatifidum during glaciations, it spread to current locations during interglaciation or postglaciation. To know the details of refuges and the migration routes, more work are needed in future survey.

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Effects of Exogenous Ethylene on Physiology and Alkaloid Accumulations in Catharanthus roseus

CHANG Bo-Wen, LIU Jie, ZHONG Peng, GUO Xiao-Rui

Bulletin of Botanical Research 2018, 38 (2): 284-291. DOI: 10.7525/j.issn.1673-5102.2018.02.017

Abstract （1001）

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Catharanthus roseus, as a medicinal plant, contains more than 100 terpenoid indole alkaloids(TIAs), including vinblastine and vincristine which focus on anti-cancer effects. In order to study the comprehensive influence of exogenous ethylene on growth, physiology, and terpenoid indole alkaloid accumulations, C.roseus seedlings were cultured as experimental material and contrasted the data of biomass, growth rate of root and stem, photosynthetic parameter and alkaloid contents with or without ethephon treatment, and the correlations between these indexes were analyzed. The ethylene release amount and the expression of erf(ethylene-response factor) gene were increased under exogenous ethylene treatment. Ethephon inhibited biomass, longitudinal growth but promoted lateral thickening growth of stem; in addition, net photosynthetic rate( P _n) and stomatal conductance( G _s) were decreased leading from non-stomatal factor. The biosynthesis of four alkaloids including strictosidine(STR), catharanthine(CAT), vindoline(VIN) and vinblastine(VINB) were accelerated in ethephon-treatment group, and the expression of key enzyme genes( str and CrPRX) were up-regulated by ethephon. By correlation analysis, we found that relationships among alkaloid contents, growth indexes and photosynthetic parameters were significant and photosynthetic parameters were significant. It were positive relation between three alkaloids including CAT, VIN, VINB and stem diameter(SD)( P < 0.05). On the contrary, there was a significantly negative relations between these three alkaloids and biomass(B), height(H), root length(RL), net photosynthetic rate( P _n)( P < 0.05). Our study will provide useful information for mechanism research of exogenous ethylene regulating TIAs biosynthesis in C.roseus.

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Simplified Genome SSR Information and Development of SSR Primers of Halenia ellipitica(Gentianaceae)

WANG Jiu-Li, CHEN Shi-Long, XING Rui, SONG Xiang-Jie, ZHU Ming-Xing, ZHANG Fa-Qi

Bulletin of Botanical Research 2018, 38 (2): 292-297. DOI: 10.7525/j.issn.1673-5102.2018.02.018

Abstract （1116）

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The restriction-site associated DNA sequencing(RAD-seq) technology was used to analyze simple sequence repeats(SSR) information of Halenia ellipitica D. Don; SSR markers for this species were developed based on RAD-seq data. We identified 6 201 SSR loci with at least 100 bp at two ends, including five types(di-nucleotide, tri-nucleotide, tetra-nucleotide, penta-nucleotide, and hexa-nucleotide). Primers of 3 865 loci were designed successfully. Among which, amount of tri-nucleotide SSR loci is the most. Repeat motif type number is 316, and type number of penta-nucleotide motif is the most(91). Afterward, 65 designed primers were randomly selected to represent the five SSR types, and tested using annealing temperature gradient PCR. Subsequently, 32 individuals from four natural populations of H. elliptica were employed to estimate the SSR primers that can be amplified. One monomorphic and 13 polymorphic SSR loci were developed according to the result of PCR and Polyacrylamide gel electrophoresis. The number of alleles of the 13 polymorphic SSR loci is high(mean 5.462), and no linkage relation was found( P<0.01); ten loci deviate from HWE( P<0.01) and have many homozygotes(observed heterozygosity mean of 0.226); the range of inbreeding coefficient is in -0.443-1(mean of 0.656); the gene flow N _m is 0.474.

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Variation in Flavonoids Biosynthesis during Seed Germination and Post-germination Growth in Astragalus membranaceus

YANG Nan, WANG Xi, GUO Xiao-Rui, LIU Yang, TANG Zhong-Hua, WANG Hong-Zheng

Bulletin of Botanical Research 2018, 38 (2): 298-305. DOI: 10.7525/j.issn.1673-5102.2018.02.019

Abstract （1272）

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Based on the stage definitions of seed germination and post-germination growth, we investigated the variations of contents of three major flavonoids during the two stages in Astragalus membranaceus and its variety of mongholicus. Besides, in order to understand how the variations were induced, the transcriptions of pathway enzyme involving in flavonoid biosynthesis were profiled. The contents of flavonoids increased during the two stages in both A.membranaceus and the variety. With the detection of gene transcriptions of the pathway enzymes, only the enzymes in upstream of the pathway contributed to the increase of flavonoids in the variety of mongholicus during the germination, whereas it was that almost all the pathway enzymes were involved in A.membranaceus. The transcriptions of pathway enzymes regulated the accumulations of flavonoids during post-germination growth, selectively with a pattern of time delay.

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Meta-analysis Identify Candidate Genes for Cold Stress Response in Arabidopsis

DUAN Ben-Yao, ZHANG Xue-Jie, FAN Shou-Jin, ZHANG Luo-Yan

Bulletin of Botanical Research 2018, 38 (2): 306-315. DOI: 10.7525/j.issn.1673-5102.2018.02.020

Abstract （1067）

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Cold stress is one of the major environmental limitations to agricultural productivity throughout the world. Overcoming the defects of microarray sequencing analyses caused by heterogeneity in individual studies and prediction of cold stress response candidate genes and pathways in Arabidopsis thaliana are requisite for cold tolerant crop-plant design and mining potential salt responses mechanisms. A literature searching strategy was used to identify salt stress response expression profiling studies in Arabidopsis from the PubMed database, the Gene Expression Omnibus(GEO) database, and the ArrayExpress database. Eight studies were filtered from the 1 166, 298 and 695 identified studies. The batch effect correction and the effect size analysis based on the NetworkAnalyst web tool were combined to integrate 8 microarray datasets, and 1 553 up-regulated and 4 327 down-regulated were identified with in low temperature treated samples. From the functional enrichment analyses results and the differently spatio-temporal expression patterns of transcriptional factors(TFs), protein kinases(PKs) and several plant hormone-related genes, plant hormone related stress signal construction pathways, ion and protein kinases related signaling pathways and transcriptional regulation were predicted to participate in regulating cold stress response in Arabidopsis.

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The in vitro Culture for Vaccinium vitis-idaea Stem Apex

LI Yan-Xia, DENG Ji-Feng, LI Gui-Jun

Bulletin of Botanical Research 2018, 38 (2): 316-320. DOI: 10.7525/j.issn.1673-5102.2018.02.021

Abstract （846）

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In order to improve the breeding rate and qualities of Vaccinium vitis-idaea seedlings, in our study, V.vitis-idaea stem apexes were used as research materials, then rapid propagation systems of plants induced by stem apex in vitro culture was established. The results indicated that the optimal induced stem apex in vitro culture is WPM(improved) +ZT 0.05 mg·L ^-1 +IBA 0.5 mg·L ^-1 +GA ₃ 0.1 mg·L ^-1. The success rate of plants induced by 0.1-0.3 mm long stem apex was only 28%, while such a rate of plants induced by 0.3-0.5 mm long stem apex was up to 68%; then stem apex reached to 0.5-0.7 mm, the success rate was 79%, and the success rate of plants induced by 0.7-1.0 mm long stem apex reached up to 94%.

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