农杆菌侵染条件下豇豆高频再生基因型的筛选

doi:10.3969/j.issn.1004-1524.2018.09.07

浙江农业学报 ›› 2018, Vol. 30 ›› Issue (9): 1489-1495.DOI: 10.3969/j.issn.1004-1524.2018.09.07

农杆菌侵染条件下豇豆高频再生基因型的筛选

周雯^{1, 2}, 吴新义², 汪宝根², 吴晓花², 鲁忠富², 汪颖², 徐沛², 李国景^{2, *}

1.浙江师范大学化学与生命科学学院,浙江金华 321000;
2.浙江省农业科学院蔬菜研究所,浙江杭州310021

收稿日期:2018-01-15 出版日期:2018-09-25 发布日期:2018-10-15
作者简介:周雯(1992—),女,安徽宣城人,硕士研究生,研究方向为植物学。E-mail: 17855871590@139.com

Screening of cowpea genotypes with high-frequency regeneration in Agrobacterium-mediated transformation

ZHOU Wen^{1, 2}, WU Xinyi², WANG Baogen², WU Xiaohua², LU Zhongfu², WANG Ying², XU Pei², LI Guojing^{2, *}

1. College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321000, China;
2. Institute of Vegetables, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2018-01-15 Online:2018-09-25 Published:2018-10-15
Contact: 李国景,E-mail: ligj@mail.zaas.ac.cn
Supported by:
“十三五”国家重点研发计划(2016YFD0100204-32); 浙江省公益性项目(LGN18C150009); 浙江省农业新品种选育重大科技专项“蔬菜新品种选育”领域(2016C02051-7-3); 国家自然科学基金(31572135,31772299); 浙江省自然科学基金(LY15C150002)

摘要/Abstract

摘要： 在农杆菌侵染条件下,对51份豇豆基因型材料进行组织培养和芽诱导分化试验。经初筛后,按再生率对不同基因型材料进行等级划分,对再生率在80%以上的10个基因型材料进行重复试验。结果表明,白八月豇的繁殖系数和不定芽诱导率均最高,分别为1.93和86.40%,该地方品种可在后续研究中作为一个较好的基因型材料用于转基因功能分析等研究。

关键词: 豇豆, 基因型, 农杆菌, 再生率

Abstract: In this study, 51 cowpea genotypes were tested for tissue culture and shoot induction in Agrobacterium-mediated transformation. After a preliminary screening for regeneration, the rate of shoot regeneration was graded and 10 genotypes with a regeneration rate above 80% were tested further in three replicates. White August Cowpea was found to show the highest reproduction coefficient and induction rate of adventitious buds, which were 1.93 and 86.40%, respectively. Therefore, this genotype may be used as a good explant source in the future transgenic studies.

Key words: cowpea, genotype, Agrobacterium tumefaciens, regeneration rate

中图分类号:

S643.4

周雯, 吴新义, 汪宝根, 吴晓花, 鲁忠富, 汪颖, 徐沛, 李国景. 农杆菌侵染条件下豇豆高频再生基因型的筛选[J]. 浙江农业学报, 2018, 30(9): 1489-1495.

ZHOU Wen, WU Xinyi, WANG Baogen, WU Xiaohua, LU Zhongfu, WANG Ying, XU Pei, LI Guojing. Screening of cowpea genotypes with high-frequency regeneration in Agrobacterium-mediated transformation[J]. , 2018, 30(9): 1489-1495.

参考文献

[1] 何礼.我国栽培豇豆的遗传多样性研究及其育种策略的探讨[D].成都:四川大学,2002.
HE L. Studies on the genetic diversity and breeding strategy of Chinese cultivated cowpea[D]. Chengdu: Sichuan University, 2002. (in Chinese with English abstract)
[2] SOLLETI S K, BAKSHI S, SAHOO L, et al. Additional virulence genes in conjunction with efficient selection scheme, and compatible culture regime enhance recovery of stable transgenic plants in cowpea via Agrobacterium tumefaciens -mediated transformation[J]. Journal of Biotechnology , 2008, 135(1): 97-104.
[3] RAVEENDAR S, IGNACIMUTHU S. Improved Agrobacterium mediated transformation in cowpea Vigna unguiculata L. Walp.[J]. Asian Journal of Plant Sciences , 2010, 9(5): 256-263.
[4] 郭兵福, 马岩松, 张丽娟,等. 不同大豆基因型对农杆菌敏感性研究及优异种质筛选[J]. 植物遗传资源学报, 2015, 16(3):569-575.
GUO B F, MA Y S, ZHANG L J, et al. Assessment of Agrobacterium sensitivity for different soybean genotypes and suitable germplasm screening[J]. Journal of Plant Genetic Resources , 2015, 16(3):569-575. (in Chinese with English abstract)
[5] 王升吉,吴元华,王洪岩.大豆不同外植体组织培养及再生研究[J]. 沈阳农业大学学报, 1999,30(3):255-259.
WANG S J, WU Y H, WANG H Y. Studies on the regeneration of tissue culture of different exophytes in soybean[J]. Journal of Shenyang Agricultural University , 1999,30(3):255-259. (in Chinese with English abstract)
[6] AKCAY U C, MAHMOUDIAN M, KAMCI H, et al. Agrobacterium tumefaciens -mediated genetic transformation of a recalcitrant grain legume, lentil ( Lens culinaris Medik)[J]. Plant Cell Reports , 2009, 28(3): 407-417.
[7] SANYAL I, SINGH A K, AMLA D V. Agrobacterium tumefaciens -mediated transformation of chickpea ( Cicer arietinum L.) using mature embryonic axes and cotyledonary nodes[J]. Indian Journal of Biotechnology , 2003, 2(4):524-532.
[8] TRIPATHI L, SINGH A K, SINGH S, et al. Optimization of regeneration and Agrobacterium -mediated transformation of immature cotyledons of chickpea ( Cicer arietinum L.)[J]. Plant Cell , Tissue and Organ Culture , 2013, 113(3): 513-527.
[9] 白守信, 梁红卫. 豌豆组织培养中的器官发生及影响分化频率的几个因素[J]. 西北植物研究, 1983,3(1):49-55.
BAI S X, LIANG H W. Several factors of organogenesis and frequency of differentiation in pea tissue culture[J]. Acta Botanica Boreali-Occidentalia Sinica , 1983, 3(1):49-55. (in Chinese with English abstract)
[10] KWAPATA K, SABZIKAR R, STICKLEN M B, et al. In vitro regeneration and morphogenesis studies in common bean[J]. Plant Cell , Tissue and Organ Culture , 2010, 100(1): 97-105.
[11] BRAR M S, ALKHAYRI J M, MORELOCK T E, et al. Genotypic response of cowpea Vigna unguiculata (L.) to in vitro regeneration from cotyledon explants[J]. In Vitro Cellular & Developmental Biology-Plant , 1999, 35(1): 8-12.
[12] RAJI A, ORIERO E, ODEYESE B, et al. Plant regeneration and Agrobacterium -mediated transformation of African cowpea ( Vigna unguiculata (L.) Walp) genotypes using embryonic axis explants[J]. Journal of Food Agriculture & Environment , 2008, 6(3/4): 350-356.
[13] GARCIA J A, HILLIE J, GOLDBACH R. Transformation of cowpea ( Vigna unguiculata ) cells with an antibiotic resistance gene using Ti-plasmid derived vectors[J]. Plant Science , 1986, 44(1):37-46.
[14] GARCIA J A, HILLIE J, GOLDBACH R. Transformation of cowpea ( Vigna unguiculata )cells with a full length DNA copy of cowpea mosaic virus mRNA[J]. Plant Science , 1987, 48(2):89-98.
[15] ILORI C O, PELLEGRINESCHI A. Transgene expression in cowpea ( Vigna unguiculata (L.) Walp.) through Agrobacterium transformation of pollen in flower buds[J]. African Journal of Biotechnology , 2011, 10(56): 11821-11828.
[16] CHAUDHURY D, MADANPOTRA S, JAIWAL R, et al. Agrobacterium tumefaciens -mediated high frequency genetic transformation of an Indian cowpea ( Vigna unguiculata L. Walp.) cultivar and transmission of transgenes into progeny[J]. Plant Science , 2007, 172(4): 692-700.
[17] DIALLO M S, NDIAYE A R, SAGNA M, et al. Plants regeneration from African cowpea variety ( Vigna unguiculata L. Walp.)[J]. African Journal of Biotechnology , 2008, 7(16):2828-2833.
[18] 罗倩. 豇豆子叶节与幼胚再生体系建立与优化[D]. 成都:四川农业大学, 2013.
LUO Q. Establish and optimize regeneration system of cowpea ( Vigna unguiculata L. walp.) cotyledonary nodes and immature embryos[D]. Chengdu: Sichuan Agricultural University, 2013.

农杆菌侵染条件下豇豆高频再生基因型的筛选

Screening of cowpea genotypes with high-frequency regeneration in Agrobacterium-mediated transformation

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