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    Screening and Phenotypic Identification of Seed Coat Pigmentation Mutants in Arabidopsis thaliana
    Na LI, Xiaonan WANG
    Bulletin of Botanical Research    2023, 43 (1): 59-68.   DOI: 10.7525/j.issn.1673-5102.2023.01.007
    Abstract3670)   HTML34)    PDF(pc) (5322KB)(913)       Save

    Flavonols play important roles in plant responses to various environmental stresses and in the regulation of seed coat development. In this study, a transparent seed coat mutant was obtained through ethyl methane sulfonate(EMS) mutagenesis screening, compared with wild-type Arabidopsis thaliana(Col-0), the mature seed color of the mutant was yellow, and its phenotypic trait was controlled by a recessive single gene. Using map-based cloning and fine-mapping technology, the mutant gene TT4(At5G13930) was mapped to BAC of MAH20 on chromosome 5, and the 1 299th base C of the TT4 was mutated to T, resulting in the 324th amino acid glycine mutated to glutamic acid. TT4(transparent testa 4) encoded a structural gene for flavonoid synthesis, chalcone synthase(CHS), and after mutation, the seed coat was transparent and the seed color was yellow, then the mutant was named tt4-1. The phenotype of brown seed coat was restored by functional complement mutants, which further demonstrated the important role of TT4 in regulating the development of seed coat color. The tissue expression analysis of TT4 was conducted by promoter coupling GUS assay, and it was found that TT4 gene was expressed in the roots, stems, leaves of plant seedlings and flowers respectively. Physiological phenotypic analysis showed that the mutant tt4-1 seeds germinated earlier, and the seedlings had shorter taproots, as well as more lateral roots and root hairs, larger stomatal opening and higher water loss rate in mature leaves compared with the wild type. This study would lay a theoretical basis for further elaborating the function of TT4 gene.

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    Proteomic Screening and RcBSK7 Resistance of Ricinus communis under Cadmium(Cd) Stress
    Huibo ZHAO, Zhiqiang ZHAO, Chunguang BAO, Qi WEN, Ruxin LI, Fenglan HUANG
    Bulletin of Botanical Research    2023, 43 (1): 36-50.   DOI: 10.7525/j.issn.1673-5102.2023.01.005
    Abstract3494)   HTML27)    PDF(pc) (8953KB)(754)       Save

    In order to reveal the mechanism of Ricinus communis response to heavy metal cadmium(Cd) stress, and screen out the resistance genes involved in Cd stress, by observing the seed germination and plant growth status, castor plants treated with water were used as controls to study their effects under three doses(300, 700 and 1 000 mg·L-1) of Cd stress. The response mechanism is expected to provide new ideas for revealing the defense and detoxification mechanism of castor plant under in Cd stress. Using differential proteomics to analyze the network regulation mechanism of castor under Cd stress, that is, with the increase of Cd stress concentration, the castor plant blocks the absorption of heavy metal Cd by roots, improves their own antioxidant capacity, and inhibits Cd2+ operation. As well as the induction of programmed cell death and other detoxification processes to resist Cd stress damage; according to the results of omics analysis, a significantly different gene RcBSK7 was screened. Through functional verification in Arabidopsis, it can be seen that this gene can improve tolerance of castor plants to Cd stress. This study enhances the understanding of the diversity and complexity of castor plants under three Cd stresses, and provides a valuable theoretical basis for the identification of Cd-tolerant genes and the restoration of heavy metal pollution in the soil.

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    Genetic Diversity and Genetic Structure of the Rare and Endangered Plant Ferula fukanensis
    Lei YANG, Qiumei CAO, Ying FENG, Wenjun LI
    Bulletin of Botanical Research    2023, 43 (1): 51-58.   DOI: 10.7525/j.issn.1673-5102.2023.01.006
    Abstract3355)   HTML24)    PDF(pc) (707KB)(1057)       Save

    To clarify the genetic diversity and genetic structure of a rare and the endangered small population of Ferula fukanensis, it is the basis and prerequisite to develop effective conservation and management strategies. The genetic diversity and structure of 87 endangered plants F. fukanensis from three populations based on 10 pairs of SSR primers with good polymorphism and stable amplification were analyzed respectively. The results showed that small population of F. fukanensis had relatively high genetic diversity, the Nei’s gene diversity index(hS) among the populations was 0.514, the total Nei’s gene diversity index(hT) was 0.516, the observed heterozygosity(Ho) was 0.881, the expected heterozygosity(He) was 0.512, the Shannon information index(I) was 0.836 and the percentage of polymorphic bands(PPB) was 100%, and the genetic differentiation degree of F. fukanensis was very low(Fst=0.007), 95.9 % of the variation of F. fukanensis occurred within populations, and there was no significant correlation between genetic distance and geographical distance, 66.7% of the populations suffered from genetic bottlenecks. The results showed that F. fukanensis had abundant genetic variation and high evolutionary potential. Based on the current situation of this wild population, it is suggested to establish a protected area, and carry out in situ conservation and strengthen ex situ conservation measures such as introduction and artificial breeding to assist the conservation of F. fukanensis. This study can provide theoretical support for the conservation of plant resources of F. fukanensis and has important theoretical and practical significance.

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    Genetic Transformation and Function Analysis of PsnHB13 and PsnHB15 of Populus simonii × Populus nigra
    Zhanmin ZHENG, Yubing SHANG, Guangbo ZHOU, Di XIAO, Yi LIU, Xiangling YOU
    Bulletin of Botanical Research    2023, 43 (3): 340-350.   DOI: 10.7525/j.issn.1673-5102.2023.03.003
    Abstract3125)   HTML10)    PDF(pc) (3156KB)(80)       Save

    Transcriptome analysis in Populus simonii × P. nigra overexpressing PsnCycD1;1 showed PsnHB3 and PsnHB15 are differential genes. Analysis of PsnHB13 and PsnHB15 conservative domains through InterPro tool. The STRING software was utilized to explore PsnHB13 and PsnHB15 proteins interaction network analysis. Yeast two hybrid was used to verify the interaction proteins,calculate the ratio of leaf length to width,the ratio of dry weight to fresh weight of leaves and stems of transgenic plants,and analyze the transcriptome of PsnHB13 overexpression plants. Conserved domain analysis showed that PsnHB13 and PsnHB15 genes belonged two subfamilies,showing that PsnHB13 mainly contains the HDZip I domain,and PsnHB15 mainly contains the HDZip Ⅲ domain. ,Each gene is screened to 10 interacting genes. Moreover,PsnHB15 has a higher probability of interaction with family proteins. Yeast two-hybrid assays demonstrated that PsnHB15 interacts with PsnHB13 and PsnHB13 interacts with PsnCycD1;1. The aspect ratio of leaves in transgenic lines overexpressing PsnHB13 and PsnHB15 increased in the early seedling stage.The ratio of dry weight to fresh weight in stems of PsnHB13 overexpression lines increased significantly(P<0.05). The analysis of the transcriptome revealed that Gene Ontologies(GO) enrichment analysis identified 3 significantly enriched GO terms,including response to chemical,response to organic substance,regulation of RNA metabolic process. KEGG pathway enrichment analysis demonstrated that transcription factors,plant hormone signal transduction and Cytochrome P450 were significantly enriched. These differential genes,including MADS-box transcription factor,MYBP transcription factor,ERF1 transcription factor,GH3 auxin responsive genes,SAUR protein family. PsnHB13 and PsnHB15 play an important role in the growth and development of Populus simonii × P. nigra,and are key genes to explore the growth law of plants and reveal the relationship between cell cycle and growth regulation.

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    Analysis on Leaf Color and Growth Variation of Transgenic BpGLK Betula pendula ‘Dalecarlica'
    Li CAO, Yunli YANG, Tianfang LI, Jing JIANG
    Bulletin of Botanical Research    2023, 43 (3): 351-360.   DOI: 10.7525/j.issn.1673-5102.2023.03.004
    Abstract3122)   HTML8)    PDF(pc) (1713KB)(121)       Save
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    Analysis of the Promoter Sequence and Response Characteristics of the BpPIN3 gene in Betula platyphylla
    Kun CHEN, Gonggui FANG, Huaizhi MU, Jing JIANG
    Bulletin of Botanical Research    2022, 42 (4): 592-601.   DOI: 10.7525/j.issn.1673-5102.2022.04.009
    Abstract2750)   HTML20)    PDF(pc) (1993KB)(194)       Save

    As an important auxin efflux carrier family in plants, PIN protein family shows extensive physiological effects in plant growth and development. In order to further understand the function of BpPIN3 and explore its response to different hormone signals and abiotic stresses during Betula platyphylla development,the BpPIN3 promoter sequence was analyzed by bioinformatics methods. Tissue expression patterns of roots, stems, leaves and apical buds of annual and biennial B. platyphylla clones were analyzed respectively. The seedlings were used as conduct hormone induction and light stress treatment with 100 μmol·L-1 auxin(IAA), 100 μmol·L-1 gibberellin(GA3), 200 μmol·L-1 abscisic acid(ABA) and long light conditions, respectively. RNA was extracted from leaves and roots of B. platyphylla at 0, 2, 4, 8, 16, 24 and 48 h after hormone treatment and at 0, 1, 3, 6, 12, 24, 48 and 72 h after light stress, and qRT-PCR was used to analyze the expression of BpPIN3. The results showed that the BpPIN3 promoter sequence contained different types of auxin response elements such as gibberellin, abscisic acid, and methyl jasmonate, as well as multiple stress-related cis-regulatory elements. BpPIN3 was expressed in many tissues of birch in different growth years, especially in leaves, and the expression of BpPIN3 in the second year was higher than that in the first year. The relative expression of BpPIN3 gene was different in different parts under different treatment conditions. IAA and GA3 could induce the up-regulation of BpPIN3 expression in leaf. Under ABA treatment, except for 16 and 48 h, the expression pattern of BpPIN3 gene was opposite to that under IAA treatment. IAA, GA3 and ABA could induce the expression of BpPIN3 in root, but shading stress induced the expression of BpPIN3 gene in leaf, the relative expression of BpPIN3 gene was significantly higher than that of the control(0 h) from 12 h in root, IAA, GA3 and ABA could induce the expression of BpPIN3 in root, shading stress induced the expression of BpPIN3 gene in leaf; the relative expression of BpPIN3 gene was significantly higher than that of the control (0 h)from 12 h in root. The results suggested that BpPIN3 gene played the important regulatory role in the growth and development of B. platyphylla,and the signal transduction pathways of IAA, GA3 and ABA and plant light response.

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    Characteristics of Complete Chloroplast Genome and Phylogenetic Analysis of Rhodiola himalensis (Crassulaceae)
    Yu ZHANG, Xu SU, Yuping LIU, Tao LIU, Changyuan ZHENG, Dandan SU, Yanan WANG, Ting LÜ
    Bulletin of Botanical Research    2022, 42 (4): 602-612.   DOI: 10.7525/j.issn.1673-5102.2022.04.010
    Abstract2250)   HTML21)    PDF(pc) (3850KB)(279)       Save

    The complete chloroplast genome of Rhodiola himalensis was sequenced, assembled and annotated using high-throughput sequencing technology, and R. himalensis was used as material, an endemic medical species to the Qinghai-Tibet Plateau. The results showed that the complete chloroplast genome of R. himalensis was 151 074 bp in length with a GC content of 37.8%. It had a typical tetrad structure with a long single copy region, a short single copy region and a pair of inverted repeat regions, and the sequence length of them was 82 309, 17 017 and 25 875 bp, respectively. The complete chloroplast genome encoded 130 genes, including 86 protein coding genes, 37 tRNA genes and seven rRNA genes. Based on the chloroplast genome of R. himalensis, a total of 25 513 codons was detected, among which the largest proportion of codons was Leu. In the IRa and IRb regions of R. himalensis, the gene of rps19 and ycf1 was deletion, and the gene of trnH in LSC region was contraction. And R. himalensis and R. sacra had the closest genetic relationship were also found. Besides, the variation frequency of single nucleotide polymorphism(SNP) was the highest within the SSC region. The complete chloroplast genome of R. himalensis was reported, assembled, annotated and analyzed, which could provide the theoretical basis for the study of genetic diversity and rational development and utilization of R. himalensis in the future.

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    Construction of Suppression Expression Vector and Genetic Transformation of BpSPL9 gene from Betula platyphylla
    Jinxia DU, Tingting SHEN, Haoran WANG, Yiping LIN, Huiyu LI, Lianfei ZHANG
    Bulletin of Botanical Research    2023, 43 (1): 30-35.   DOI: 10.7525/j.issn.1673-5102.2023.01.004
    Abstract2176)   HTML29)    PDF(pc) (1428KB)(760)       Save

    To clarify the function of SPL gene in growth and development of Betula platyphylla,the BpSPL9 gene was cloned, and the expression vector of BpSPL9 was constructed by CREST technology, and the genetic transformation was carried out by Agrobacterium mediated method, and the phenotype of the transgenic lines was observed, respectively. The results showed that the transgenic lines with BpSPL9 suppressed were successfully obtained. The height of transgenic lines was significantly lower than that of non-transgenic lines, with shorter node spacing and smaller leaf area. The contents of 6-BA and IAA in transgenic lines were lower than those of the non-transgenic lines. It is speculated that BpSPL9 gene participated in plant growth and development by affecting the synthesis of auxin and cell division.

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    Establishment and Application of Genetic Transformation System for Populus simonii× P. nigra ‘Baicheng’
    Xu HE, Yuan GAO, Qunye ZHANG, Chenguang ZHOU, Wei LI, Shuang LI
    Bulletin of Botanical Research    2023, 43 (5): 667-678.   DOI: 10.7525/j.issn.1673-5102.2023.05.004
    Abstract1540)   HTML11)    PDF(pc) (4918KB)(191)       Save

    Stems of Populus simonii×P. nigra ‘Baicheng’ in vitro plants were selected as explants and MS medium was used to establish tissue culture system by adjusting the hormone concentrations of 6-BA, NAA, TDZ and IBA. Based on the tissue culture system, the optimal concentration of kanamycin and infection time were confirmed to establish Agrobacterium tumefaciens-mediated genetic transformation system for P. simonii×P. nigra ‘Baicheng’. By using this system, the transgenic plants overexpressed a key tension wood formation regulator LBD39 (Lateral Organ Boundaries Domain) were created successfully. The results showed that the tissue culture system consisted of three stages, including adventitious bud differentiation induction (MS+ 0.5 mg·L-1 6-BA+0.1 mg·L-1 NAA+0.001 mg·L-1 TDZ, shoot differentiation rate=92.6%), stem induction (MS+0.2 mg·L-1 6-BA+0.05 mg·L-1 NAA, multiplication coefficient=6.5) and rooting induction (1/2 MS+0.4 mg·L-1 IBA, rooting percentage=100%). The optimal kanamycin concentration for genetic transformation was 30 mg·L-1 and the optimal infection time was 20 min, and the transgenic plants were obtained successfully after 30 d of adventitious bud differentiation induction, 15-30 d of stem induction and 25 d of rooting induction with 2% transformation efficiency respectively. By using this system, five overexpressed plants of LBD39 were obtained, and the transformation efficiency was 3.3%.

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    Evaluation on Waterlogging Tolerance of Different Tomato Germplasm
    Luhua ZHOU, Junyi FANG, Zimo XIONG, Weifeng WU, Jiarui LIU, Qiao LU, Hongqing LING, Danyu KONG
    Bulletin of Botanical Research    2023, 43 (5): 657-666.   DOI: 10.7525/j.issn.1673-5102.2023.05.003
    Abstract1476)   HTML13)    PDF(pc) (4406KB)(169)       Save

    To screen out the tomato(Solanum) germplasm with waterlogging tolerance, 50 tomato germplasm were selected as the research object, and the survival rate and physiological indexes were compared after semi-submerged and submerged treatments respectively. The results showed that 12 waterlogging tolerant tomato germplasm with 100% survival rate after seven days submergence treatment were selected, and 12 waterlogging-tolerant germplasm were observed and distributed in S. lycopersicumS. lycopersicum var. cerasiforme and S. pimpinellifolium respectively. Further analysis on waterlogging-tolerant tomato TK017 and waterlogging-sensitive tomato TK039 revealed that tomato TK017 could control the content of reactive oxygen species more effectively under submergence condition, and improved the waterlogging tolerance of tomato.

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    Comparative Proteomics on Leaves of Triadica sebifera Clones under Salt Stress
    Dezong SUI, Baosong WANG
    Bulletin of Botanical Research    2023, 43 (5): 679-689.   DOI: 10.7525/j.issn.1673-5102.2023.05.005
    Abstract1335)   HTML8)    PDF(pc) (2944KB)(167)       Save

    In order to investigate the molecular mechanisms of protein responses in Triadica sebifera to salt stress, two T. sebifera lines, including salt-sensitive P18(SS18) and salt-tolerant P21(ST21) were selected and treated with 0.4% NaCl solution. Isotope tags for relative and absolute quantitation(iTRAQ) were used to quantify leaf proteins abundance changes at different stress periods(0, 24, 72 h). A total of 279 and 106 differentially expressed proteins(DEPs) were detected in SS18 and ST21, respectively. Four proteins, including catalase, extension factor 1-α, H+-ATPase c domain protein and thioredoxin, were significantly up-regulated in two clones under salt stress, suggesting that they may be important potential target proteins in the salt-tolerant response of T. sebifera’s. Pathway enrichment analysis showed that the DEPs of T. sebifera leaves under salt stress were primarily associated with photosynthesis, carbohydrate metabolism, amino acid metabolism, and fatty acid metabolism. The protein-protein interaction network showed that 5 core proteins(24 h) and 3 core proteins (72 h) involved in glucose decomposition and energy metabolism in SS18, while ST21 had 5 (24 h) and 4 (72 h) core proteins involved in carbon metabolism, photosynthesis, carbon fixation, photosynthesis and related to chlorophyll a-b binding. ST21 leaves improved metabolic pathways of sugars, amino acids and fatty acids, and accumulated a large number of small molecular soluble substances such as soluble sugars, amino acids and organic acids under salt stress, which might be an important aspect of salt stress response.

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    Establishment of an Efficient Regeneration System in Iris lacteal var. chinensis with Immature Embryo
    Wenjie AN, Yinjie WANG, Qingquan LIU, Yongheng YANG, Ting ZHANG, Shaoru FAN, Yongxia ZHANG, Haiyan YUAN
    Bulletin of Botanical Research    2023, 43 (3): 333-339.   DOI: 10.7525/j.issn.1673-5102.2023.03.002
    Abstract1216)   HTML10)    PDF(pc) (2035KB)(123)       Save

    The effect of sampling time and plant growth regulators on the induction of callus, somatic embryo and adventitious bud were investigated with immature embryos of Iris lacteal var. chinensis as explants. The results showed that the most feasible sampling time for immature embryos was 35-45 d after pollination, the young embryos were fully developed, but the endosperm was not completely hardened, and the young embryo was easy to dissect. The optimum medium for callus induction was MS+1.0 mg·L-1 2,4-D+0.2 mg·L-1 6-BA with the induction rate of 83.33%. The appropriate medium for somatic cell induction was MS+0.25 mg·L-1 2,4-D+0.05 mg·L-1 6-BA with the induction rate of 71.67%. The most suitable medium for adventitious bud induction and proliferation was MS+0.2 mg·L-1 TDZ+0.05 mg·L-1 NAA with the induction rate of 78.33% and proliferation coefficient 3.65 in this experiment. The rooting rate was more than 95% after four weeks cultivation on MS medium supplemented with 0.1 mg·L-1 NAA. Then, the medium was washed off and explants were transplanted to sterilized vermiculite, and the survival rate was more than 95%. This study has established an efficient regeneration system of I. lacteal var. chinensis embryos, which can lay a foundation for genetic transformation and gene function analysis of I. lacteal var. chinensis.

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    Fruiting and Seed Characteristics of Abies in Northwest Yunnan
    Dan WANG, Zhongshuai ZHANG, Qingyin ZENG, Xuemin HAN
    Bulletin of Botanical Research    2023, 43 (5): 647-656.   DOI: 10.7525/j.issn.1673-5102.2023.05.002
    Abstract828)   HTML16)    PDF(pc) (1495KB)(205)       Save

    To explore the fruiting characteristics and seed characteristics and their variation patterns of Abies species distributed in Northwest Yunnan, and to provide material support and theoretical basis for promoting the regeneration and restoration of Abies forest, the specimens and cones of seven Abies species distributed in Northwest Yunnan were collected and identified, and the seed morphological characteristics, seed viability, and fruiting characteristics were analyzed, and then the differences among species and regions were explored separately. The results showed that the length, width and thickness of seeds of seven Abies species in Northwest Yunnan were 6.04-10.22 mm, 2.03-3.32 mm and 1.26-2.24 mm, respectively, and the thousand-grain weight was 4.26-30.50 g. The thousand-grain weight was significantly correlated with seed length, width and thickness (r>0.8, P<0.01). The average rates of plump, empty and insect-eaten seeds of seven Abies species in Northwest Yunnan were 27.51%, 4.92% and 67.58%, respectively. The rate of plump seed was low, and that of empty and insect-eaten seeds was high. Abies ernestii var. salouenensis was not further analyzed in this article due to field investigations revealing that most of them had no cone and only one tree’s cones were collected in this study. For the other six species, the rate of insect-eaten seeds did not differ significantly between species(P=0.750), or between regions(P=0.204). However, there were interspecies differences in the rate of plump seeds(P=0.005) and empty seeds(P=0.007). There were interspecific(P=0.008) and interregional(P=0.036) differences in Abies seed viability. Therefore, seed viability of Abies was affected by both species characteristics and environmental factors.

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    Effects of Abiotic Stresses on the Intracellular and Extracellular ATP Levels of Tobacco Suspension Cells
    Li LI, Xin WANG, Yuejing ZHANG, Lingyun JIA, Hailong PANG, Hanqing FENG
    Bulletin of Botanical Research    2023, 43 (2): 179-185.   DOI: 10.7525/j.issn.1673-5102.2023.02.003
    Abstract493)   HTML27)    PDF(pc) (1080KB)(852)       Save

    In this study, the effects of NaCl, PEG(6000) and low temperature stresses on the levels of iATP (intracellular ATP) and eATP(extracellular ATP) of tobacco suspension cells BY-2(Nicotiana tabacum ‘Bright Yellow-2’) were studied respectively. The results showed that the tobacco suspension cells treated with 50-200 mmol·L-1 NaCl significantly increased membrane permeability(P<0.05), the levels of iATP and eATP decreased significantly at 100 and 200 mmol·L-1 NaCl stresses(P<0.05). With the increase of PEG mass concentration(50, 100 and 200 g·L-1), the membrane permeability and eATP level increased. Under 200 g·L-1 PEG stress, eATP level increased significantly to 3.4-fold of the control(P<0.05), iATP level decreased significantly to 0.5-fold of the control(P<0.05). After the treatment with low temperature at 0-10 ℃, the membrane permeability and iATP level increased to different extents. iATP level increased to 1.9-fold of the control at 0 ℃, while eATP level decreased significantly to 0.8-fold the control at 10 ℃(P<0.05). The above results indicate that iATP is the source of eATP in plant cells, iATP level is not the only factor that affects eATP level under abiotic stresses. The way in which plant cells transport ATP and other factors may affect eATP level under abiotic stresses.

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    Genomic Survey Analysis of Endangered Plant Tetraena mongolica
    Ziyin WANG, Bingru LIU, Zihao LI, Jifei WANG, Pengbin YANG
    Bulletin of Botanical Research    2023, 43 (2): 186-193.   DOI: 10.7525/j.issn.1673-5102.2023.02.004
    Abstract464)   HTML23)    PDF(pc) (1760KB)(98)       Save

    To assess the size and complexity of the genome of the endangered plant Tetraenamongolica, and reveal the hyper-arid mechanism of T.mongolica and further explore its characteristic genetic resources, and to better decipher the whole genome information of T.mongolica, the second-generation high-throughput sequencing technology of genome survey analysis was used as the method, and bioinformatics method was used to estimate the genome information such as heterozygosity, repeat sequence and GC content of T.mongolica respectively. The results showed that the genome size of T. mongolica was 1 079.25 Mb, the corrected genome size was 1 065.84 Mb, and the heterozygosity rate was 0.76%, the percentage of repetitive sequences was 75.25%, and the GC content was 33.57% respectively. After the initial assembly of T. mongolica genome 3 502 126 contigs were obtained, totaling 682 Mb, with an N50 of 187 bp. It was hypothesized that T. mongolica genome belonged to a homo-tetraploid complex genome, which was difficult to assemble by whole-genome sequencing. Due to the high heterozygosity of T. mongolica, the third-generation high-throughput sequencing technology(single-molecule sequencing) combined with chromatin region capture technology could be used subsequently, which was expected to finally obtain a high-quality whole-genome map of T. mongolica.

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    Genome‑wide Identification and Stress Response Analysis of Cyclic Nucleotide-gated Channels( CNGC) Gene Family in Populus trichocarpa
    Xueying WANG, Ruiqi WANG, Yang ZHANG, Cong LIU, Dean XIA, Zhigang WEI
    Bulletin of Botanical Research    2022, 42 (4): 613-625.   DOI: 10.7525/j.issn.1673-5102.2022.04.011
    Abstract372)   HTML22)    PDF(pc) (7555KB)(274)       Save

    The plant cyclic nucleotide-gated channels(CNGC) family has a variety of biological functions and plays an important role in the growth and development of plants and the response to stress. In this study, bioinformatics methods and qRT-PCR were used to analyze the basic physicochemical properties, structural characteristics, phylogeny, gene structure and conserved motif, cis-acting elements of the promoter and gene expression patterns of the PtrCNGC gene family members respectively. The results showed that a total of 19 PtrCNGC genes were identified in the whole genome of Populus trichocarpa. PtrCNGC gene family could be divided into four subgroups(Ⅰ, Ⅱ, Ⅲ and Ⅳ), and subgroup Ⅳ could be divided into two subgroups(Ⅳa and Ⅳb). The proteins encoded by the PtrCNGC genes were all basic proteins, and only one member of the PtrCNGC gene family was hydrophobic protein, while all the other members were hydrophilic proteins. A total of 19 PtrCNGCs were distributed unevenly on 11 chromosomes of P. trichocarpa, while the remaining 8 chromosomes had no members distributed. The PtrCNGC gene family contained seven pairs of homologous genes and the Ka/Ks value between them was far less than one. There was little difference in gene structure and protein conserved motif distribution among the family members. Predictive analysis of cis-acting elements in the promoter region of the PtrCNGC gene sequence showed that there were acting elements related to various hormones and stress. The qRT-PCR results showed that the expression of PtrCNGC gene family was specific in different tissues, with higher expression levels in stems and lower expression levels in roots and leaves. Under salt stress and drought stress, most members of the same branch of the PtrCNGC gene family showed similar expression patterns. The results of this study provid a reference for further study on the function of PtrCNGC family in abiotic stress.

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    Prokaryotic Expression, Preparation Polyclonal Antibody and Protein Expression of BoMAPK4 of Ornamental Kale( Brassica oleracea var. acephala)
    Heming LI, Hongtao QIN, Deqiang WEI, Xu LI, Xingguo LAN
    Bulletin of Botanical Research    2022, 42 (4): 584-591.   DOI: 10.7525/j.issn.1673-5102.2022.04.008
    Abstract246)   HTML19)    PDF(pc) (4214KB)(201)       Save

    The mitogen-activated protein kinase 4(MAPK4)cDNA were isolated from the stigma of ornamental kale(Brassica oleracea var. acephala)self-incompatibility line(S13-bS13-b )by RT-PCR. The obtained BoMAPK4 cDNA contained a 1 122 bp open reading frame in length, encoded 373 amino acids, and had a serine/threonine domain, no signal peptide and transmembrane structure. The deduced amino acid sequence of BoMAPK4 shared 99.7%, 99.5% and 95.4% identity with Brassica napus BnMAPK4, Brassica rapa BrMAPK4, Arabidopsis thaliana AtMAPK4, respectively. The ORF of BoMAPK4 was constructed into the prokaryotic expression vector pET-14b and transformed into Escherichia coli BL21(DE3)for expression. SDS-PAGE showed that BoMAPK4 protein was specifically expressed at a molecular weight of 45 kDa. The recombinant BoMAPK4 was obtained by affinity chromatography, and the polyclonal antibody against BoMAPK4 was prepared by immunizing mice. The total proteins of sepals, petals, anthers, stigmas, styles and ovaries of ornamental kale were extracted, and protein expression was detected by Western blot. The results showed that BoMAPK4 was less expressed in petals, anthers and ovary, more expressed in sepals and styles, and the highest expressed in stigmas. The study lays the foundation for exploring the biological functions of BoMAPK4.

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    Comparison of Genetic Diversity Between Wild and ex- situ Conservation Populations of Cupressus chengiana
    Ermei CHANG, Jianfeng LIU, Yuening HUANG, Hongli LI, Bingyan SHAN, Zeping JIANG, Xiulian ZHAO
    Bulletin of Botanical Research    2022, 42 (5): 772-779.   DOI: 10.7525/j.issn.1673-5102.2022.05.008
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    To evaluate the genetic diversity level and provide theoretical basis of ex-situ conservation populations of Cupressus chengiana, the genetic diversity, genetic structure, and gene flow of ex-situ conservation populations and wild populations of C. chengiana were studied respectively. The principal component analysis(PCA), cluster analysis, construction of molecular evolutionary tree, genetic diversity and genetic structure of three wild populations and ex-situ conservation populations based on SNP locus were obtained respectively from Genotyping-by-Sequencing technology(GBS) sequencing. The results showed that high quality clean data 118 321 514 728 bp was obtained by GBS sequencing, and 1 947 047 tags were developed. After being screened, a total of 1 259 610 of high-quality SNP sites were retained. The valley value of cross validation error rate determined that the optimal number of clusters was 1. The phylogenetic tree indicated that most of individuals ex-situ conservation populations and wild populations were clustered together. Both of combined ex-situ conservation populations and wild populations of C. chengiana had higher genetic diversity, and their value of heterozygosity(Ho), expected heterozygosity(He), Shannon information index(Shi), inbreeding coefficient(Fis), and percentage of polymorphic loci(Pic) were 0.181 5-0.272 0, 0.223 2-0.300 3, 0.331 0-0.464 9, 0.178 0-0.246 5 and 0.272 2-0.309 2, respectively. Genetic diversity of ex-situ conservation cultivated populations was relatively higher than the wild populations of C. chengiana, the value of He and Shi was 0.300 3 and 0.464 9, respectively. The wild populations of C. chengiana of Baiwan tunnel(BW)_vs_Songgang town(SA) had the greater genetic differentiation and the gene flow(Nm) were lower than other populations(Fst=0.091,Nm=2.496). The results indicated that genetic differentiation had not occurred among these C. chengiana populations, and the gene had exchanges frequently among the ex-situ conservation populations and wild populations of C. chengianaFst<0.05, Nm>4). There was a close genetic relationship between the ex-situ conservation individuals and the wild populations individuals, and there was no obvious clustering phenomenon. Thus, transplanting endangered plants was a better method in ex-situ conservation. This paper would provide a reference for the ex-situ conservation populations of wild populations C. chengiana and provided a theoretical reference for the conservation of other tree germplasm resources.

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    Genetic Diversity Analysis from Different Geographic Populations of Linaria vulgaris var. sinensis by rpl32- trnL Fragment
    Yujie ZHANG, Pengxia SHI, Fan ZHAO, Xiaodan CHEN, Kun SUN, Hui ZHANG
    Bulletin of Botanical Research    2022, 42 (5): 780-789.   DOI: 10.7525/j.issn.1673-5102.2022.05.009
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    In order to investigate the population genetic characteristics of Linaria vulgaris, four different geographic populations containing 62 individuals were analyzed by using rpl32-trnL fragment of chloroplast DNA. The results showed that there were 15 haplotypes and 76 loci existing in 4 populations. The total genetic diversity of L. vulgaris reached a high level(Hd=0.878, π=0.003 88, K=2.994). The AMOVA analysis showed that the genetic variations were mainly maintained within the populations(51.49%), which was consistent with the genetic characteristics of self-incompatibility in L. vulgaris. The genetic differentiation among four geographic populations was significant(FST=0.466 14), while the gene flow between different geographic populations was rather low(Nm=0.29). The genetic differentiation between different populations was positively correlated with the geographical distance but no significance(R2=0.36, P>0.05). The results of neutrality test showed that the Heshui population(HS) might experience some bottleneck events during its evolution (Fu and Li’s D=-2.450 49,P<0.05), and the evolutionary feature of the other populations was consistent with the neutral hypothesis. This study revealed that the reproductive strategies, geographic isolation, and habitat disturbance were not only responsible for the rather high level of genetic diversity in the populations distributed in the northern area of China as well as their genetic structure characteristics, but also it might provide a scientific basis for the further study of conservation genetics in L. vulgaris.

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    Effect of Fruit-Expander on Cone Performance of Pinus sylvestris var . mongolica
    Yujin XU, Xihe WANG, Pingyang LI, Yulei LI, Dexiao LI, Yunyang WU, Quan SUN, Lili GE, Xiyang ZHAO
    Bulletin of Botanical Research    2022, 42 (5): 762-771.   DOI: 10.7525/j.issn.1673-5102.2022.05.007
    Abstract168)   HTML17)    PDF(pc) (2021KB)(132)       Save

    Mongolian pine(Pinus sylvestris var. mongolica) is an important timber, landscaping and afforestation tree species in North China. In order to provide theoretical basis for high quality, high yields and stable yields of cones and putting forward scientific management strategies, the effect of fruit-expander treatment on cones and nuts characters of P. sylvestris var. mongolica was studied. P. sylvestris planted in Qinhe seed orchards, Wangqing County, Yanbian, Jilin Province were used as materials to compare and analyze the effects of traits, including cone yield, cone quality, seed yield and seed quality, treated with two fruit-expanders including chlorinated urea and α-sodium-naphthyl acetate respectively. The average seed traits were higher than those of the control, indicating that fruit-expander treatment could promote the seed growth of P. sylvestris var. mongolica. The cone weight, cone yield, seed long and seed width of P. sylvestris var.mongolica were all significant in different variation sources(P<0.001), indicating that the fruit-expander treatment had a significant effect on the cone quality and yield. The phenotypic variation coefficient of cone yield and seed weight reached to 28.54%, 33.88%, respectively. The cone traits were comprehensively evaluated for each treatment, and the optimal treatment was 10 mg·L-1 Chlorinated urea, and the real gains of cone length, cone width and cone weight were 11.81%, 4.65% and 24.35%, respectively. The optimal treatment according to the seed traits was 10 mg·L-1α-sodium-naphthynl acetate, and the real gains of seed yield per cone, seed weight, seed long, seed width, wing long and wing width were 21.32%, 24.15%, 0.24%, 9.16%, 0.33% and 2.16%, respectively. The results showed that chlorinated urea(10 mg·L-1) and α-sodium-naphthyl(10 mg·L-1) treatments could significantly improve the cone size and seed bearing of P. sylvestris var. mongolica.

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