Table of Content

15 March 2016, Volume 36 Issue 2

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Metapilea,a New Genus of Urticaceae from Guangxi

WANG Wen-Tsai

2016, 36(2):  164-166.  doi:10.7525/j.issn.1673-5102.2016.02.002

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A genus belonging to the tribe Lecantheae Wedd. of the family Urticaceae, Metapilea, is described as new from Guangxi Autonomous Region. This genus is closely related to Pilea Lindl., and from the latter differs in its piatillate capitate inflorescences with oblong discoid receptacles and with numerous bracts and pistillate flowers borne on the adaxial surface of receptacle. Besides, it is also related to Lecanthus Wedd., and from the latter differ in its sessile pistillate capitula with conduplicate oblong receptacles and lacking involucres and in its neither cucullate nor corniculate pistillate tepals. Without doubt, these two advanced genera, Metapilea and Lecanthus, have both originated from the primitive genus Pilea.

Spore Propagation of Four Ferns

ZHU Xiao-Feng1,2；SHEN Hui1*；JIN Dong-Mei1；ZHAO Guo-Hua1,3； LIU Bao-Dong3；LING Tie-Jun2；YAN Yue-Hong1

2016, 36(2):  167-176.  doi:10.7525/j.issn.1673-5102.2016.02.003

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The development of fern resources is attracting more attention in recent years, however, current spore reproductive technology is immature, and has become a bottleneck. We cultured the spores of four typical ferns, including Blechnum orientale, Matteuccia struthiopteris, Ceratopteris thalictroides and Adiantum malesianum, observed spore propagation characteristics and time of the gametophyte development at each stage. By compiling the reported data, we analyzed the culture conditions, such as light and temperature, in four key development stages, including spore germination, and the formation of prothalli, sexual organs, and juvenile sporophyte. Longer illumination time may promote spore germination. Higher light intensity may prolong the formation of prothallus. Growth rate and temperature had no correlation during gametophyte formation in 20-25℃. Our research is useful for establishing the system of fern propagation and may promote the exploration of fern resources.

Optimization of Leaf Regeneration and Genetic Transformation System of Populus tomentosa

LI Chun-Li；WANG Xiao-Jing；DING Qiang-Qiang；WU Han*

2016, 36(2):  177-183.  doi:10.7525/j.issn.1673-5102.2016.02.004

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The experiment was conducted to optimize leaf regeneration and genetic transformation of Populus tomentosa, and performed to select best leaf regenerative method, rooting medium, with an emphasis on hormone combination and compositions, and to select best explants and Agrobacterium concentration. We demonstrated that twenty days were needed for generation of cluster buds through direct differentiation of leaf tissues. With the mixed hormones of 6-BA and TDZ, we observed the 100% leaf differentiation. Further, the propagation coefficient of single leaf was improved by the step of shoot elongation. We also drastically improved root initiation and growth if gelrite was used when compared to commonly used agar, judging with the number and the length of the roots, and the shoots can root for about eight days. For genetic transformation efficiencies, our experiment showed that leaf explants after numerous subcultures can be infected by Agrobacterium but the transformation efficiency was reduced, if the primary cultured leaves were selected as explants and the concentration of bacteria liquid was OD₆₀₀=0.3, transgenic callus formation was greater than 83.5%, all shoots survived the antibiotic selection were 100% transgenic, and the transformation efficiency was more than 55%. We report a highly efficient genetic transformation method for P.tomentosa, it is significantly different from the previously published methods. The regeneration of leaf discs through direct differentiation, the mixed use of hormone 6-BA and TDZ, and the step of shoot elongation can improve the efficiency of leaf tissue culture, also using gelrite as the coagulant in rooting medium without the addition of hormone promotes the initiation and growth of roots, and improves the plant growth as well. The primary cultured leaves are more suitable for genetic transformation of P.tomentosa.

Main Anthocyanin Profiles in Petals of Freesia hybrida

XU Yi-Qian1；YUAN Yuan1；TAO Xiu-Hua2；YANG Juan3；SHI Yi-Min1；TANG Dong-Qin1*

2016, 36(2):  184-189.  doi:10.7525/j.issn.1673-5102.2016.02.005

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We analyzed the anthocyanin profiles in 6 cultivars of freesia(Freesia hybrida). The petal color was observed according to the Royal Horticultural Society Color Card(RHSCC). Petal pigment type was identified by characteristic color reaction. UPLC-PAD and UPLC-Q-TOF-MS were used for qualitative and quantitative analysis of anthocyanins. The flavonoids in petals of all cultivars were detected, while carotenoids were not detected; meanwhile, every cultivar, except for ‘Shangnong Ruxiang’, contained anthocyanins in petals. Among five anthocyanincontained cultivars, the highest anthocyanin content was observed in ‘Shangnong Purple Rose’, followed by ‘Shangnong Chenghong’. Totally 6 anthocyanin components were detected in petals of these cultivars, including Delphinidin 3,5-O-glucoside, Petunidin 3,5-O-glucoside, Delphinidin 3-O-glucoside, Malvidin 3,5-O-glucoside, Petunidin 3-O-glucoside and Malvidin 3-O-glucoside. Three anthocyanin components were detected in petals of ‘Shangnong Zixueqing’ and ‘Shangnong Gongfen’, of which were majorly malvidin and petunidin derivated anthocyanins. Malvidin-anthocyaninswith single or double glucosides was present in petals of ‘Shangnong Zixueqing’, while all anthocyanins in petals of ‘Shangnong Gongfen’ bonded with double glucosides, such as Delphinidin 3,5-O-glucoside, Petunidin 3,5-O-glucoside and Malvidin 3,5-O-glucoside, major Petunidin 3-O-glucoside and minor Delphinidin 3-O-glucoside in petals of ‘Shangnong Purple Rose’. While only one anthocyanin component was detected in petals of ‘Shangnong Golden Queen’ and ‘Shangnong Chenghong’, which was Delphinidin 3,5-O-glucoside and Malvidin 3,5-O-glucoside, respectively.

Adventitious Bud Induction with Immature Embryo of Herbaceous Peony

WEI Dong-Xia1,2,3；GAO Kai4；YUAN Yan-Bo1；TANG Zheng-Jiao1；YU Xiao-Nan1,2,3*

2016, 36(2):  190-194.  doi:10.7525/j.issn.1673-5102.2016.02.006

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Culture of zygotic embryos is an effective way to accelerate the breeding process of herbaceous peony. With the zygotic embryos of three herbaceous peony(Paeonia lactiflora Pall.) cultivars(‘Fen Yu Nu’, ‘Zhu Sha Pan’ and ‘Tuan Ye Hong’), we studied the affection of different embryo age, explant types, sterilization methods and starting medium formula on root initiation in vitro. The suitable development period were 50 and 70 d after flowering for embryo-culture of ‘Zhu Sha Pan’ and ‘Tuan Ye Hong’, respectively. We got better results when using excised zygotic embryos rather than keeping part of endosperm reserved. The optimal disinfection method for explant was that firstly treating carpel for 30-60 s with 70% alcohol, then for 20 min with 2% NaClO, and finally for 10 min with 2% NaClO. The optimal initial medium was 1/2MS+1.0 mg·L^-1 IAA+0.5 mg·L^-1 GA₃.

Effects of Soil Fauna on C Dynamics in Different Sized Forest Gaps in Pinus massoniana Plantation

LI Xun1；LIU Yang1,2,3；ZHANG Yan1；LIU Hua1；YANG Wan-Qin1,2,3；ZHANG Jian1,2,3*

2016, 36(2):  195-203.  doi:10.7525/j.issn.1673-5102.2016.02.007

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Soil fauna has an important effect on litter nutrient release, and the effect could vary with litter types and be controlled by environmental factors such as temperature and precipitation. We to characterized the effect of soil fauna on C dynamics in different sized forest gaps (G1: 100 m², G2: 400 m², G3: 900 m², G4: 1 600 m²) during litter decomposition of a Pinus massoniana plantation in a hilly area of the upper reaches of Yangtze River. A field litterbag experiment was conducted in P.massoniana plantation of Sichuan, China, from January 2014 to November 2014. Samples of airdried leaf litter for P.massoniana(Pm) and Cinnamomum camphora(Cc) were placed in nylon litterbags (20 cm×20 cm, 10 g per bag) of two different mesh sizes (3.00 and 0.04 mm). The amount of C released were investigated in different sized forest gaps during the first year of decomposition, i.e., Spring, Summer, Autumn and Winter. There was a significant difference in the amount of litter C released through soil fauna among different sized gaps: medium sized gap (G3: 900 m²) promoted the C release of Cc and Pm. The content of C displayed a declining trend in Spring and Summer, but with an obvious enrichment process in Winter and Autumn. Soil fauna imposed a positive effect on C release, the contribution of soil fauna to the C loss rate displayed as Cc>Pm. Based on the above results, soil fauna imposed a positive effect on C release in P.massoniana plantation. When we reconstruct the low-function forest, we can take advantage of medium sized gap and introduce native broadleaf tree species Cc which is easier to be decomposed by soil fauna to accelerate nutrient cycling and maintains the soil fertility of P.massoniana plantation.

Effects of Water Extraction from Roots of Carya illinoensis on Seed Germination and Seedling Growth of Two Crops

ZHANG Quan1；YAO Xiao-Hua1*；TENG Jian-Hua2；SHAO Wei-Zhong3；FU Song-Ling4

2016, 36(2):  204-210.  doi:10.7525/j.issn.1673-5102.2016.02.008

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In order to probe the impact of water extraction from roots of Carya illinoensis for Triticum aestivum L. and Glycine max(L.) Merr, we studied the effects of different levels(0.025, 0.05 and 0.1 g·mL^-1) of water extraction from roots of C.illinoensis on the seeds and seedling growth of T.aestivum L. and G.max(L.) Merr with indoor bio-assay. The water extract prominently inhibited the seed germination and seedling growth of T.aestivum L. and G.max(L.) Merr with concentration effect(weak inhibition in low concentration, high inhibition in high concentration). The descending order of inhibition of water extract for germination rate, germination energy, germination index and vigor index of T.aestivum and G.max(L.) Merr were vigor index, germination energy, germination index, germination rate, and vigor index, germination energy, germination rate, germination index, respectively. The descending order of allelopathic effects of water extract on shoot height, root length, fresh weight and dry weight of T.aestivum and G.max(L.) Merr were both root length, shoot height, fresh weight, dry weight. The water extract prominently decreased the seedling root length, but had different impacts on the dry and fresh weight of receptor due to the different species. The order of the synthesis effects of water extraction from roots of C.illinoensis on two crops is T.aestivum L.>G.max(L.) Merr. The activity of SOD in the 7 d seedings was decreased, but the content of MDA, the activity of POD and CAT were increased with the increasing of concentration. Comprehensively, the inhibition of the water extraction from roots of C.illinoensis is higher for T.aestivum L. than for G.max(L.) Merr.

Determining the Main Organic Compounds in Ammodendron bifolium Seeds

ZHU Yan-Lei1,2；AN Deng-Di2；SHE Xiao-Ping1*

2016, 36(2):  211-215.  doi:10.7525/j.issn.1673-5102.2016.02.009

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Ammodendron bifolium(Pall.) Yakovl. is a rare psammophyte distributed in Xinjiang of China. In order to study the adaptation of the species to desert environment, the contents of main organic compounds in its seeds were determined. Carbohydrate, soluble protein and crude fat accounted for 16.8%, 23.7% and 15.5% of seed dry weight respectively. Soluble carbohydrate accounted for 10.1% of seed dry weight, and sucrose accounted for 43.4% of soluble carbohydrate content. Contents of amino acids in proteins and free amino acids accounted for 26.5% and 0.4% of seed dry weight, respectively. Unsaturated fatty acid was more than 90% of total fat content, and oleic acid and linoleic acid were its main component.

Effect of Shade Treatment in Summer on the Expression of Genes Related to Theanine Biosynthesis in Tea Plants(Camellia sinensis)

CHEN Qi；YU Shu-Wei；JIANG Xue-Mei；ZHAO Ying；MENG Xiang-Yu；WAN Xiao-Chun*

2016, 36(2):  216-223.  doi:10.7525/j.issn.1673-5102.2016.02.010

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The shade can effectively improve the quality of tea(Camellia sinensis) by causing an increase in the concentration of theanine, the main compound that contributes to the fresh and brisk taste of tea leaves. We analyzed the influence of shade on theanine biosynthesis in relation to the expression of genes related to theanine synthesis in tea leaves during summer. Our data reveal that shade has a notable effect on the level of both theanine and total free amino acids in young leaves. The results of qPCR and western blot analysis suggest that these effects are mainly brought about via an increase in the mRNA expression of TS and promotion of TS protein synthesis. The qPCR results also suggest that the mRNA expression of all the genes related to the theanine pathway in the leaves, including glutamine synthetase(GS), glutamate dehydrogenase(GDH), glutamine 2-oxoglutarate amino-transferase(GOGAT), nitrite reductase(NiR), arginine decarboxylase(ADC), are significantly affected by shade. However, from two different expression patterns, the GS/GOGAT pathway is triggered by shade and members of this pathway show higher expression in old leaves, which reassimilate more NH⁺₄ to supply more amides to young leaves. The gene expression of NiR and ADC was markedly lower in both the old and young leaves from the shaded plants, which suggests that nitric nitrogen metabolism might be inhibited by shade treatment. Thus, shade treatment can increase the content of free amino acids especially theanine in tea leaves and improve the effective utilization of summer tea.

Effects of Complex Saline-alkali Stress on Seed Germination and Stress-resistance Physiological Characteristics of Naked Oat Seedlings

LIU Jian-Xin1,2；WANG Jin-Cheng1,2；WANG Rui-Jian1,2；JIA Hai-Yan1,2

2016, 36(2):  224-231.  doi:10.7525/j.issn.1673-5102.2016.02.011

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We studied the effects of salt-alkaline mixed stress on seeds germination, seedlings growth and leaf physiological characteristics of stress-resistance for the physiological response of a new naked oat(Avena nude) cultivar ‘Dingyou No 6’ to complex saline-alkali stress. A.nude seeds and seedlings were cultivated in sands with different complex saline-alkali solution addition according to soil salinity composition of the planting areas. Germination rate, germination index, vigor index of seeds, water content and dry weight of seedlings were significantly decreased, while the contents of superoxide anion, hydrogen peroxide and malondialdehyde and relative plasma membrane permeability in leaves were increased evidently with increasing of the complex saline-alkaline stress. The complex saline-alkaline stress increased significantly Na⁺ contents, but reduced K⁺ contents and K⁺/Na⁺. The activities of plasma membrane H⁺-ATPase, peroxidase, ascorbate peroxidase and soluble protein content in leaves of the seedlings exhibited a humped-curve, while superoxide dismutase activities and the contents of glutathione, organic acid and proline were significantly increased. Catalase activity and ascorbic acid content were decreased evidently, free amino acid content first decreased and then increased, and soluble sugar content demonstrated a rose at first, and then dropped at last rose again. The imbalance of active oxygen metabolism and the disturbance of Na⁺ and K⁺ homeostasis in leaves resulted in the inhibition of naked oat seedlings growth, and the organic acid and proline are two possible osmotic adjusting substances of naked oats that are adapted to complex salt-alkaline stress.

Sequence Analysis of Nuclear DNA  JRD5680 for Determining Genetic Diversity and Genetic Structure Analysis of Common Walnut(Juglans regia L.)

ZHANG Tian；WANG Ma-Li；ZHAO Peng*

2016, 36(2):  232-241.  doi:10.7525/j.issn.1673-5102.2016.02.012

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We studied the genetic diversity and population genetic structure of 265 individuals from different regions of China by using the roles of phenylalanine ammonia-lyase(PAL) nuclear gene JRD5680 DNA sequence of 40 common walnut(Juglans regia) and 5 out group plant species. The length of JRD5680 sequence was 809 bp, the content of G+C was 46.2% with 74 information sites. A total of 30 haplotypes, haplotype diversity variation showed low genetic diversity(H_d=0.370, π=0.005 3). There is a clear geographical structure(N_ST>G_ST), which is a regional distribution pattern. The spatial genetic structure analysis showed that the correlation between geographical isolation and genetic distance was significant(r=0.263 2; P=0.032 4*), and there was obvious geographical isolation effect among different geographic populations. The results of comprehensive mismatch analysis and neutral test can be deduced from the recent history of the different regions of the population expansion event. From the genetic structure, the walnut population can be divided into two groups(southwest and other regions). By AMOVA, the genetic variation among the populations was mainly stored in the populations(56.54%), and the genetic differentiation among the populations was higher(F_ST=0.885). The total of 5 species of Juglandaceae could be a single branch, and the JRD5680 sequence might be with high molecular identification radio in the members of the Juglandaceae.

Allelic Variation in 4-coumarate:CoA ligase(4CL) Associated with Wood Properties of Larix olgensis

WANG Yan-Hong1,2；JIA Qing-Bin1；ZHANG Lei1；ZHANG Han-Guo1*

2016, 36(2):  242-251.  doi:10.7525/j.issn.1673-5102.2016.02.013

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We used 10 provenances of Larix olgensis to study the effects of polymorphism of 4CL gene on wood quality and growth traits in L.olgensis. The polymorphism in 4CL gene was identified using DNA sequencing, 64 mean SNPs were identified,the median SNP frequency was one site per 31 bp,and the average nucleotide diversity for the sequenced regions was calculated to be π_T =0.015 77 and θ_W =0.008 78. The diversity level of synonymous nucleotide sbustituons(π_syn=0.022 56) was 1.4 of nonsynoymous nucleotide substitutions(π_nonsyn=0.015 75), suggesting that the gene misht be evolved under purifying selection at the synonoymous sites of the coding region in L.olgensis. The association between the 27 common SNPs(frequency>10%), haplotypes and wood quality and growth traits were analyzed respectively, indicating that 7 SNPs and 6 haplotypes were significantly associated with wood quality and growth traits, explaining between 0.09%-2.08% of the phenotypic variance. Ourstudy provided an important genetic foundation for molecular marker-assistant selection breeding programs with the goals of improving the wood quality and growth products in L.olgensis.

Sequence and Expression Analysis of MYB Family Genes in Betula platyphylla Suk.

LIU Hui-Zi；SUN Dan；YU Ying；ZHANG Nan；WANG Chao*

2016, 36(2):  252-257.  doi:10.7525/j.issn.1673-5102.2016.02.014

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The MYB transcription factor family is one of the important plant transcription factor family members, and they have the key function on the growth, development, cell wall biosynthesis and stress response in plant. We obtained the total of 17MYB genes, and by phylogenetic analysis, 17 MYB genes of birch belong to different subfamilies and groups. Among these genes, 10 MYBs belong to the 1R/4R subfamilies, and 7 MYBs belong to the 2R subfamily. BplMYB13 is homologous to the AtMYB46 which functions in the secondary cell wall biosynthesis. BplMYB15 and BplMYB26 belong to the group which involved in the stress response. BplMYB23 is homologous to the AtMYBs involved in the glucosinolate biosynthesis. BplMYB9, 21 and 22 have homologous with the AtMYBs functioned in phenylpropanoid biosynthesis. The expression patterns analysis among the development stages of the growth season and the tension wood and opposite wood bended for 6 h, and the normal wood were performed by real time RT-PCR. 17 MYB genes had the peak express levels from the middle of May to the middle of July, and the BplMYB13 maintained the high express levels among all the growth season in the cambium and xylem. The BplMYB13 may play the role in the cambium and xylem development. After 6 h bending, 14 MYBs were up-regulated in the tension wood compare with the opposite wood or normal wood, and 7 MYBs were increased expressed in the opposite wood than those in the normal wood. Therefore, these genes response to the artificial bending and play key roles in the physiology change during the xylem development response to the bending stress.

Cloning and expression analysis of GrCMS gene in Gentiana rigescens

ZHANG Xiao-Dong1；LI Cai-Xia1；WANG Yuan-Zhong2*

2016, 36(2):  258-265.  doi:10.7525/j.issn.1673-5102.2016.02.015

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2-C-methyl-D-erythritol 4-phosphatecytidyltransferase(CMS, EC 2.7.7.60) is the third enzyme in methylerythritol phosphate(MEP) pathway. The Open Reading Frame(ORF) of GrCMS gene was cloned by RT-PCR technology from young leaves of Gentiana rigescens based on the transcriptome of G.rigescens and its prokaryotic and the tissue specific expression analysis were performed. The ORF of GrCMS gene(accession number: KJ917164) was 933 bp long coding for a protein of 310 amino acids, and the predicted relative molecular weight of GrCMS was 34.23 kD with its theoretical pI of 7.68. The results of GrCMS protein analysis showed that GrCMS which possessed the conserved domains of CMS proteins and may localize in chloroplast was a hydrophilic stable protein without signal peptide, and it was composed of mainly α-helix(26.45%) and random coils(54.84%). By phylogenetic analysis, GrCMS was close to CrCMS of Catharanthus roseus. By prokaryotic analysis, the recombinant protein of GrCMS gene in E.coli was approximately 60.23 kD(containing GST tag protein 26 kD), which was consistent with the anticipated size. By real-time PCR analysis, GrCMS gene was primarily expressed in leaf. Our results will provide reference for further functional researches of GrCMS gene and the biosynthetic pathway of gentiopicroside.

Response of Germination on Ten Chenopodiaceae Seeds to Drought Stress and Resistances Evaluation in Tarim Basin

HAN Zhan-Jiang；CHENG Long；LI Zhi-Jun*

2016, 36(2):  266-273.  doi:10.7525/j.issn.1673-5102.2016.02.016

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The experiments were conducted with ten Chenopodiaceae seeds as materials to examine responses of seed germination and evaluate their tolerance under drought stress by PEG-6000. The final germination rates and radicle length of seeds were inhibited while the mean germination times extended continuously in various degrees with the drought intensifying in 10 Chenopodiaceae species. Seeds of Suaeda glauca, Suaeda rigida, Suaeda heterophylla, Halocnermum strobilaceum, Halostachys caspica, Salicornia europaea and Halogeton arachnoideus obtained their germination recoveries after relieving drought stresses. The limit values of drought tolerance at seed germination stage from large to small were H.caspica, H.arachnoideus, S.heterophylla, S.rigida, S.europaea, Suaeda arcuata, H.strobilaceum, Suaeda stellatiflora, Chenopodium aristatum and S.glauca. Comprehensive evaluation of drought resistances of seed germination were conducted with fuzzy subordinate function analysis by selecting the final genmination rate, mean generation time, radicle length and germination recovery rate. The drought resistances at seed germination stage from large to small were H.caspica, H.arachnoideus, S.rigida, H.strobilaceum, S.heterophylla, S.stellatiflora, S.europaea, C.aristatum, S.glauca and S.arcuata. Seed germinations of H.caspica, H.arachnoideus, S.rigida and H.strobilaceum showed the best resistances under drought stresses between ten Chenopodiaceae species, and their weighted average values of subordinate function were 0.915, 0.793, 0.762 and 0.737, respectively.

Influence of Different Vegetation Types on the Soil Inorganic Nitrogen at Small Watershed in Northern Zhejiang Province

WANG Qing-Bing1；WU Hao1；ZHANG Jian-Feng1*；CHEN Guang-Cai1；LI Ze-Bo1；WANG Li1,2；YANG Quan-Quan1

2016, 36(2):  274-282.  doi:10.7525/j.issn.1673-5102.2016.02.017

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We studied the contents of inorganic nitrogen in surface and subsurface soil within six different vegetation types at Fushi Reservoir watershed in Anji County of northern Zhejiang Province, and the influence of different vegetation types on soil inorganic nitrogen distribution. There is significant difference(P＜0.05) between the contents of soil NO^-₃-N and NH⁺₄-N at Tianjia mountain and Rihui mountain watershed, and the maximum differences of soil NO^-₃-N and NH⁺₄-N are 10.09 and 11.45 mg·kg^-1 in plant non-growing season with more drastic fluctuations when compared to the plant growing season(9.6 and 2.72 mg·kg^-1). Soil organic nitrogen accounts for more than 97.0% of the total nitrogen at six vegetation types in two seasons. By the correlation analysis, there is a close relationship between NO^-₃-N and NH⁺₄-N of 0-20 cm soil with the physical and chemical properties of soil itself for six vegetation types. The total nitrogen input to reservoir in the plant nongrowing season is higher than that in the growing season, and that of Rihui mountain watershed is significantly greater than that of Tianjia mountain watershed, 1.49 times and 1.55 times more than that of Tianjia mountain watershed in two plant seasons, respectively. The plant non-growing season shows a greater risk of nitrogen loss in this region, and it is also the key stage for the nitrogen pollution control, Metasequoia glyptostroboides forest and Echinochloa crusgalli(L.) Beauv show a larger capacity for storing and retaining nitrogen, which are suitable for large-scale construction to protect the water quality of Fushi Reservoir watershed.

Construction of Plant Expression Vector and Genetic Transformation Analysis of Arabidopsis thaliana CYCD2;1 Gene in Nicotiana tabacum

DAI Li-Juan；ZHENG Tang-Chun；LIU Cai-Xia；LI Kai-Long；QU Guan-Zheng*

2016, 36(2):  283-290.  doi:10.7525/j.issn.1673-5102.2016.02.018

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Cell division is one of the basic characteristics of organisms, which plays an extremely important role in the plant growth and development. CYCD2;1, as a regulatory factor, has been identified in regulation of cell cycle. In our study, CYCD2;1 was amplified from Arabidopsis inflorescences cDNA by PCR. Plant overexpression vector (pROKII-AtCYCD2;1) was constructed and transformed into wild-type tobacco by Agrobacterium-mediated leaf disc transformation. The transgenic plants were confirmed by PCR, and the result showed AtCYCD2;1 gene was integrated into tobacco genome. The result of qRT-PCR detection of T2 generation plants showed the mRNA of AtCYCD2;1 was transcribed in transgenic tobaccos. There were significant differences in the flower between the transgenic and wild type plant, such as the width of the corolla, the lengths of the petal and sepal, and increased volume of seed pod, which suggesting overexpression of AtCYCD2;1 has an obvious influence on the development of flowers.

Characteristic of Molecular Evolution of Notopterygium incisum Based on nrDNAITS and cpDNA rpl20-rps12 Sequence Analysis

YANG Lu-Cun1,2；LIU He-Chun1,3；ZHOU Xue-Li4；XU Wen-Hua1,2；ZHOU Guo-Ying1,2*

2016, 36(2):  291-296.  doi:10.7525/j.issn.1673-5102.2016.02.019

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We analyzed the differences between the nrDNAITS and cpDNA rpl20-rps12 sequences in Notopterygium incisum by PCR direct sequencing. Total DNA was extracted from silica-dried leaves of N.incisum using modified CTAB method. With the extracted DNA as template, nrDNA ITS and cpDNA rpl20-rps12 regions were amplified, then purified and sequenced. The length of nrDNA ITS sequence of N.incisum was 635 bp, of which 17 were variable sites with a percentage of 2.68%, the(G+C) content was 57.83%. The length of cpDNA rpl20-rps12 sequence of N.incisum was 767 bp, of which 35 was variable site with a percentage of 4.56%, the(G+C) content was 33.06%. The nrDNAITS region of N.incisum was more conserved and evolved more slowly than the cpDNA rpl20-rps12sequence. The present distribution range of N.incisum experienced range expansion by the haplotype analysis of this species, which consisted with the conclusion resulting from cpDNA genome. Therefore, the nrDNA ITS sequence of rpl20-rps12 was fit to the phylogeographic study of this species.

Genetic Relationships of 39 Cerasus Cultivars by ISSR Analysis

LIN Li；WANG Zhi-Long*；FU Tao；LIN Le-Jing

2016, 36(2):  297-304.  doi:10.7525/j.issn.1673-5102.2016.02.020

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Cerasus trees are famous ornamental plants in the world with a large number of cultivars, but the origin of most Cerasus cultivars are poorly known, resulting in some difficulties in cultivars classification. We analyzed the genetic relationships among 39 cultivars of Cerasus by intersimple sequence repeat(ISSR) molecular-marked technique, and screened 14 ISSR primers to assess the genomes of 39 cultivars of Cerasus. The result showed that a total of 109 DNA bands were amplified and 102 of which(93.58%) were polymorphic. The Nei’s genetic similarities of 39 cultivars ranged from 0.493 to 0.942 with the average of 0.727, suggesting that the genetic similarities among the 39 cultivars were relatively close. According to the Nei’s genetic similarity of 0.697, by UPGMA method cluster analysis, 39 cultivarss were classified into 2 cluster groups, and the group Ⅱ were classified into 4 subcluster groups with the genetic similarity of 0.738. The result not only supported the classification method of Kawasaki Tetsuya which classified Cerasus cultivars into 7 groups according their origin, but also showed that ISSR technique was applied to classify Cerasus cultivars. From cluster results, flower inflorescence, form and colour were important indicators for Cerasus cultivars classification.

Age Variations in Productivity and Family Selection of Open-Pollinated Families of Korean Pine(Pinus koraiensis)

ZHANG Zhen；ZHANG Han-Guo*；ZHANG Lei

2016, 36(2):  305-309.  doi:10.7525/j.issn.1673-5102.2016.02.021

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With 79 open-pollinated families of Korean pine (Pinus koraiensis) collected from plus trees of Qingshan seed orchard, we tested the growth capability of 21, 25 and 27 years at Weihe Forestry Bureau of Shangzhi City, in Heilongjiang Province. The volume of wood of growth capability was the biggest at half rotation period. Coefficient of variation was the biggest in volume of wood and the averages of coefficient of variation were 13.12%, 22.39% and 46.34% in height, diameter at breast height and volume of wood for three phases of old, respectively. The results showed highly significant family effects for growth capability at 21, 25 and 27 years. The genetic variations were under middle-level genetic control, the family heritability all was apparently higher than individual heritability for three phases of old. We selected 16 superior families and 17 superior individuals according to the volume and 20% rate of selection. The averages in the volume of superior families were 0.036 8, 0.048 6 and 0.061 6 m³ for three phases, respectively, and all were 28.67%, 15.02% and 14.07% more than that of CK, respectively. The genetic gain for individual volume were in 20.25%, 34.37% in 17 superior individuals.

FTIR Spectra and Their Taxonomic Significance of Nine Euphorbia Species in Shaanxi

TIAN Lan-Ting；ZHAO Xue-Yan；WANG Ning-Guo；LI Wei；PENG Yong；CAI Xia*

2016, 36(2):  310-315.  doi:10.7525/j.issn.1673-5102.2016.02.022

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We measured infrared spectra of nine raw herbs of Euphorbia L. distributed in Shaanxi Province by Fourier Transform Infrared(FTIR) spectrometer, and analyzed the secondary derivative infrared spectra in 1 800-400  cm^-1. By principal component analysis of the second derivative spectra, the first two principal components accounted for 77.04%, which reflected the most information of the original spectrum and could be used for identifying nine Euphorbia species. The scattered score distributive diagram and cluster analysis based on the first two principal components showed that nine raw herbs of Euphorbia L. can be divided into three classes, E.kansui was one class, E.lathylris was the other class, and E.humifusa, E.lunulata, E.pekinensis, E.hylonama, E.jolkini, E.esula and E.helioscopia fell into another class. Our result is similar to that by morphological classification, and FTIR spectroscopy combined with principal component analysis and cluster analysis could provide evidence for the taxonomic study of Euphorbia.