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Volume 35 Issue 12
Dec.  2020
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Article Contents
LEI Y, XIE Q, CHEN T, et al. Yeast Two-hybrid cDNA Library Constructed from Vitis davidii Föex Pericarps Infected by Grape Ripe Rot Pathogen, Colletotrichum viniferum [J]. Fujian Journal of Agricultural Sciences,2020,35(12):1330−1335 doi: 10.19303/j.issn.1008-0384.2020.12.006
Citation: LEI Y, XIE Q, CHEN T, et al. Yeast Two-hybrid cDNA Library Constructed from Vitis davidii Föex Pericarps Infected by Grape Ripe Rot Pathogen, Colletotrichum viniferum [J]. Fujian Journal of Agricultural Sciences,2020,35(12):1330−1335 doi: 10.19303/j.issn.1008-0384.2020.12.006

Yeast Two-hybrid cDNA Library Constructed from Vitis davidii Fex Pericarps Infected by Grape Ripe Rot Pathogen, Colletotrichum viniferum

doi: 10.19303/j.issn.1008-0384.2020.12.006
  • Received Date: 2020-11-12
  • Rev Recd Date: 2020-11-29
  • Available Online: 2020-12-10
  • Publish Date: 2020-12-31
  •   Objective  A yeast two-hybrid cDNA library on Colletotrichum viniferum-infected pericarps of the wild spine grape (Vitis davidii Föex), a cultivar highly resistant to the serious grape ripe rot in southern China, was constructed to facilitate studies on the molecular mechanism of the disease resistance in the plants.   Method  The total RNA was extracted from the pericarps of V. davidii accession, Fu'an, 1, 3, and 7d after C. viniferum inoculation and reverse-transcribed into cDNA using the SMART method. The cDNA was purified by an assay kit into a double stranded cDNA to be digested. Then, the short fragments were removed to obtain a high-quality cDNA for the subsequent cloning into a pGADT7 3-frame plasmid vector and purifying for the establishment of a primary cDNA library. After amplification, the plasmid was extracted and transformed into Yeast Y187 to produce the amplified library for final identification.  Result  The primary cDNA library had a capacity of approximately 5.2×106 cfu with a recombination rate of approximately 97.92% and the inserted fragments of good polymorphism in the lengths ranging from 400 to 2 000 bp. The harvested Y187 yeast library titer was about 6.0×107 cfu·mL−1.   Conclusion  The constructed cDNA library from C. viniferum-infected grape pericarps was adequate for yeast two-hybrid screening. It would materially aid the studies on the interacting proteins in grape plants infected by the pathogen.
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  • [1]
    OIV. 2019 Statistical report on world vitiviniculture [R/OL]. (2019-10-15) [2020-11-25]. http//www.oiv.intpublicmedias6782oiv-2019-statistical-report-on-world-vitiviniculture.pdf.
    [2]
    贺普超, 任治邦. 我国葡萄属野生种对炭疽病抗性的研究 [J]. 果树科学, 1990, 7(1):7−12.

    HE P C, REN Z B. Study on the resistances of wild Vitis species native to China to grape ripe rot [J]. Joural of Fruit Science, 1990, 7(1): 7−12.(in Chinese)
    [3]
    SUZAKI K. Improved method to induce sporulation of Colletotrichum gloeosporioides, causal fungus of grape ripe rot [J]. Journal of General Plant Pathology, 2011, 77(2): 81−84. doi: 10.1007/s10327-011-0296-z
    [4]
    赵奎华. 葡萄病虫害原色图鉴[M]. 北京: 中国农业出版社, 2006.
    [5]
    YAN J Y, JAYAWARDENA M M R S, GOONASEKARA I D, et al. Diverse species of Colletotrichum associated with grapevine anthracnose in China [J]. Fungal Diversity, 2015, 71(1): 233−246. doi: 10.1007/s13225-014-0310-9
    [6]
    LEI Y, TANG X B, JAYAWARDENA R S, et al. Identification and characterization of Colletotrichum species causing grape ripe rot in southern China [J]. Mycosphere, 2016, 7(8): 1177−1191. doi: 10.5943/mycosphere/si/2c/8
    [7]
    OO M M, OH S K. Identification and characterization of new record of grape ripe rot disease caused by Colletotrichum viniferum in Korea [J]. Mycobiology, 2017, 45(4): 421−425. doi: 10.5941/MYCO.2017.45.4.421
    [8]
    王跃进, 徐炎, 张剑侠, 等. 中国野生葡萄果实抗炭疽病基因的RAPD标记 [J]. 中国农业科学, 2002, 35(5):536−540. doi: 10.3321/j.issn:0578-1752.2002.05.014

    WANG Y J, XU Y, ZAHNG J X, et al. Identification of RAPD markers linked to ripe rot resistant gene in wild grapes native to China [J]. Scientia Agricultura Sinica, 2002, 35(5): 536−540.(in Chinese) doi: 10.3321/j.issn:0578-1752.2002.05.014
    [9]
    方中达. 植病研究方法[M]. 第3版. 北京: 中国农业出版社, 1998.
    [10]
    雷龑, 林雄杰, 陈婷, 等. 福建葡萄炭疽病病原鉴定及致病性分析 [J]. 果树学报, 2014, 31(6):1123−1127, 1199.

    LEI Y, LIN X J, CHEN T, et al. Pathogen identification and pathogenicity analysis of grape ripe rot in Fujian [J]. Journal of Fruit Science, 2014, 31(6): 1123−1127, 1199.(in Chinese)
    [11]
    FIELDS S, SONG O K. A novel genetic system to detect protein-protein interactions [J]. Nature, 1989, 340(6230): 245−246. doi: 10.1038/340245a0
    [12]
    程萍, 冯仁军, 袁克华, 等. 香蕉红素氧还蛋白酵母双杂交cDNA文库的构建及鉴定 [J]. 生命科学研究, 2009, 13(4):349−353.

    CHENG P, FENG R J, YUAN K H, et al. Construction and identification of yeast two-hybrid cDNA library of Musa acuminata rubredoxin (MaRD) [J]. Life Science Research, 2009, 13(4): 349−353.(in Chinese)
    [13]
    苏玲, 李彬, 王青, 等. 金柑花蕾酵母双杂交cDNA文库构建及评价 [J]. 基因组学与应用生物学, 2019, 38(7):3169−3173.

    SU L, LI B, WANG Q, et al. Construction and evaluation of yeast two hybrid cDNA library of floral buds from Rongan kumquat [J]. Genomics and Applied Biology, 2019, 38(7): 3169−3173.(in Chinese)
    [14]
    刘露露, 曲俊杰, 郭泽西, 等. 霜霉菌侵染后葡萄叶片酵母双杂交cDNA文库构建 [J]. 南方农业学报, 2020, 51(4):829−835.

    LIU L L, QU J J, GUO Z X, et al. Construction of a yeast two-hybrid cDNA library from Vitis vinifera leaves infected by downy mildew [J]. Journal of Southern Agriculture, 2020, 51(4): 829−835.(in Chinese)
    [15]
    姜赟, 史自航, 付欣, 等. 番茄花柄脱落过程的酵母双杂交cDNA文库构建 [J]. 沈阳农业大学学报, 2018, 49(2):129−135.

    JIANG Y, SHI Z H, FU X, et al. The Construction of a yeast two-hybrid cDNA library of tomato pedicel during abscission. [J]. Journal of Shenyang Agricultural University, 2018, 49(2): 129−135.(in Chinese)
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