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Molecular Detection and Resistance Analysis of Soybean-lines with HrpZPsta(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2020年02期
Page:
220-226
Research Field:
Publishing date:

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Title:
Molecular Detection and Resistance Analysis of Soybean-lines with HrpZPsta
Author(s):
JI Cai-feng;  SHAO Rui-chao;  ZHANG Qi;  ZHOU Jing-xuan;  LIU Tian-yi;  GUO Xuan-lin;  ZHANG Liang;  WANG Pi-wu
(College of Agronomy, Jilin Agricultural University, Changchun 130118, China)
Keywords:
Soybean;  JN27-119-21;  Phytophthora root rot;  hrpZPsta Gene;  Disease resistance
PACS:
-
DOI:
10.11861/j.issn.1000-9841.2020.02.0220
Abstract:
In order to verify the genetic stability of hrpZPsta gene in transgenic line JN27-119-21 and detect the disease resistance of the progenies of JN27-119-21, so as to provide a theoretical reference for breeding new soybean varieties resistant to Phytophthora root rot, in this experiment, T7 and T8 generation of soybean-lines JN27-119-21 with hrpZPsta gene were used as test materials for identificating the disease resistance ability by PCR and Southern Blot molecular biological detection and hypocotyl infection to identify the resistance to Phytophthora of the offspring of JN27-119-21. The results showed: The exogenous hrpZPsta gene can be stably inherited in high-generation transgenic lines, the hrpZPsta gene was successfully integrated into the genome of soybean receptor Jinong 27 with single copy, and the integration sites were different. The hrpZPsta gene was expressed in the roots, stems and leaves of the transformed lines, the average relative expression of the T7 was 2.877, 1.336 and 7.734, the average relative expression of T8 was 3.612, 1.746 and 8.627, and the relative expression level the T7 and T8 were both leaf>root>stem. The offspring of the transgenic lines were resistant to the disease resistance level of Phytophthora root rot, while the recipient strain was medium resistant and the disease resistance was significantly improved.

References:

［1］张君,王丕武,关淑艳,等.高产高油中晚熟广适性春大豆新品种吉农27号的选育［J］.种子, 2010,29(4): 106-107. (Zhang J, Wang P W, Guan S Y. Breeding of a new spring soybean variety Jinong 27 with high yield and high oil in late maturity［J］.Seed,2010,29(4):106-107.)［2］周恩远,刘丽君,祖伟,等.春大豆农艺性状与品质相关关系的研究［J］.东北农业大学学报,2008,39(2):145-149. (Zhou E Y, Liu L J, Zu W, et al. Study on the relationship between agronomic traits and quality of spring soybean［J］.Journal of Northeast Agricultural University, 2008,39(2):145-149.)［3］Pace P F, Weaver D B. Additional genes for resistance to frogeye leaf spot race 5 in soybean［J］.Crop Science,1993,33(6):1144-1145.［4］许丽,李玥莹,林凤. 植物抗病的分子基础与研究进展［J］. 杂粮作物,2006, 26(6): 428-432. (Xu L, Li M Y, Lin F. Molecular basis and research progress of plant disease resistance［J］. Rain Fed Crop, 2006, 26(6):428-432.)［5］Canaday C H, Schmitthenner A F. Effects of chloride and ammonium salts on the incidence of phytophthora root and stem rot of soybean［J］. Plant Disease,2010,94(6):758-765.［6］Schmitthenner A F. Problems and progress in control of phytophthora root rot of soybean［J］.Plant Disease,1985,69: 362-368.［7］姜兆远,高洁,张佳环.烟草野火病原菌hrpZpsta基因的克隆与表达［C］//第四届中国植物细菌病害学术研讨会论文集. 杭州：中国植物病理学会植物病原细菌专业委员会，2008:68-72. (Jiang Z Y, Gao J, Zhang J H.Clone and expression of hrpZpsta genes form Pseudomonas syringae pv.tabaci［C］//The 4th China National Symposium on Phytobacteriology. Hangzhou: Professional Committee of Plant Pathogenic Bacteria, Plant Pathology, 2008:68-72.)［8］He S Y, Huang C H,Collmer A. Pseudomonas syringae pv. syringae harpinpss; Protein that is secreted via the hrp pathway and elicts the hypersensitive response in plant［J］. Cell,1993,73:1255-1266．［9］Taguchi F,Tanaka R,Kinoshita S, et al.Harpinpsta from pseudomonas syringae pv. tabaci is defective and deficient in its expression and HR-inducing activity［J］. Journal of General Plant Pathology, 2001, 67: 116-123.［10］Lin H J, Cheng H Y, Chen C H. Plant amphipathec proteins delay the hypersensitive response caused by harpinpss and pseudomonas syringae pv. syringae［J］. Physialogical and Molecular Plant Pathology, 1997, 51: 367-375.［11］Strobel N E, Ji C, Goplan S, et al. Induction of systemic acquired resistance in cucumber by pseudomonas syringae pv. syringae 61 hrpZpss protein［J］. The Plant Journal,1996,9:431-439.［12］Lindgren P B. The role of hrp genes during plant-bacterial inter-actions［J］.Annual Review of Phytopathology,1997, 35:129-152．［13］Wei Z M, Beer S V. Harpin from erwinia amylovora induces plant resistance［J］. Acta Horticulture,1996, 411: 223-225.［14］余榕捷,洪岸,庞义,等. SZZ-Harpin 融合蛋白在苏云金芽孢杆菌中的表达及活性测定［J］.微生物学通报,2003,30(2):11-15. (Yu R J,Hong A,Pang Y,et al. Expression and biological assay of SZZ-Harpin fusion protein in bacillus thuringiensis［J］. Microbiology,2003,30(2):11-15.)［15］傅华欣,陆云梅,毛华方,等.康壮素在塑料大棚草莓上的应用效果［J］.安徽农业科学,1999,27(6):596. (Fu H X,Lu Y M,Mao H F,et al.Studies on the technique of applying Harpin for the diseases control of strawberry grown in greenhouse［J］.Journal of Anhui Agricultural Sciences,1999,27(6):596.)［16］高正良,马国胜,周本国,等.康壮素对烟叶产质量及综合抗病虫能力的影响［J］.烟草科技,1999(5):43-44. (Gao Z L,Ma G S,Zhou B G,et al. Effect of Harpin on yield, quality and disease, insect resistance of flue-cured tobacco［J］. Tobacco Science and Technology,1999(5):43-44.)［17］Cooper B. A defective movement protein of TMV in transgenic plants confers resistance to multipleviruss whereas the functional analog increases susceptibility［J］.Virology,1995,206: 307-313.［18］王春艳, 关淑艳, 王丕武, 等. 农杆菌介导的BADH—CMO基因转化玉米愈伤组织的初步研究［J］.吉林农业大学学报,2012,34(5): 503-507, 510. (Wang C Y, Guan S Y,Wang P W, et al. Preliminary studies on transforming bivalent salt-tolerant gene into maize callus by agrobacterium tumefaciens［J］. Journal of Jilin Agricultural University,2012,34(5): 503-507, 510.)［19］张林.新抗虫基因C〖STBX〗ry1AB13-2〖STBZ〗的克隆及其在大豆中的遗传转化和功能鉴定［D］.长春: 吉林农业大学, 2018:9. (Zhang L. Cloning of new insect resistant gene CryAB13-2 and its genetic transformation and functional identification in soybean［D］. Changchun: Jilin Agricultural University,2018: 9.)［20］魏桂芳. 大豆转Harpin(Xooc)蛋白编码基因(〖STBX〗hrf2〖STBZ〗)及其在后代中的遗传表达研究［D］.南京: 南京农业大学, 2014. (Wei G F. The inheritance and expression of 〖STBX〗HRF2〖STBZ〗 gene encoding harpinxoocin subsequent generations of transgenic soybean［D］. Nanjing: Nanjing Agricultural University, 2014.)［21］袁瑞玲,孟小林,徐进平,等.HrpZ激发对几种植物防御相关酶活性的影响［J］.安徽农业科学,2008,36(32):13951-13953,14066. (Yuan R L,Meng X L,Xu J P, et al. Effects of hrpZ excitation on activity of defense-related enzymes in several plant［J］. Anhui Agricultural Sciences, 2008,36(32):13951-13953, 14066.)［22］张佳环,李娟,高洁.大豆细菌性斑点病菌〖STBX〗hrpZPsg12〖STBZ〗基因增强病原菌对大豆的致病性并引起烟草过敏性反应［J］.安徽农业科学,2011,39(15):9009-9012. (Zhang J H,Li J,Gao J. 〖STBX〗hrpZPsg12〖STBZ〗 gene of pseudomonas syringae pv.glycinea can enhance pathogenicity of the pathogenon soybean and cause the hypersensitive response of tobacco［J］. Anhui Agricultural Sciences,2011,39(15): 9009-9012.)［23］任秀艳. Harpins蛋白在黄瓜上的研究进展［J］.安徽农业科学,2010,38(22):11714-11716.(Ren X Y.Progress research on cucumber by harpin［J］.Anhui Agricultural Sciences,2010,38(22):11714-11716.)［24］韩青梅,孟颢光,曹丽华.过敏素类蛋白研究进展［J］.河南农业大学学报,2004, 38(3):319-322.(Han Q M, Meng H G, Cao L H. Progress of studies on harpins［J］.Journal of Henan Agricultural University,2004, 38(3):319-322.)［25］廖俊杰.采用CTAB法快速提取植物DNA［J］.天津农业科学,1993(3):26.(Liao C J. Rapid extraction of plant DNA by CTAB method［J］.Tianjin Agricultural Science,1993(3):26.)［26］魏洪波,王丕武,张卓,等.大豆查尔酮还原酶GmCHR基因的克隆及其在烟草中的表达［J］.吉林农业大学学报,2014,36(5): 546-553.(Wei H B,Wang P W,Zhang Z,et al.Cloning of chalcone reductase GmCHR gene and its expression in tobacco［J］.Journal of Jilin Agricultural University,2014,36(5):546-553.)［27］王春艳,关淑艳,王丕武,等.农杆菌介导的BADH—CMO基因转化玉米愈伤组织的初步研究［J］.吉林农业大学学报,2012,34(5):503-507,510.(Wang C Y,Guan S Y,Wang P W,et al.Preliminary studies on transforming bivalent salt-tolerant gene into maize callus by agrobacterium tumefaciens［J］.Journal of Jilin Agricultural University,2012,34(5):503-507,510.)［28］李琦,宋阳,于淼,等.转hrpZpsta和chi双价广谱抗病基因大豆对疫霉根腐病的抗性分析［J］.吉林农业大学学报,2019,41(1):11-16.（Li Q,Song Y,Yu M,et al.Resistance analysis of hrpZpsta and chi bivalent broad-spectrum disease resistance soybean to Phytophthora root rot［J］.Journal of Jilin Agricultural University,2019,41(1):11-16.）［29］尹俊琦,王楠,周莹,等.两个大豆品种转hrpZpsta基因后代对大豆灰斑病的抗性分析［J］.大豆科学, 2013,32(2): 238-241. (Yin J Q,Wang N, Zhou Y, et al. Analysis on resistance to frogeye leaf spot between two species of transgenic soybean progenies with hrpZpsta gene［J］. Soybean Science, 2013,32(2):238-241.)［30］谷晓娜,刘振库,王丕武,等. hrpZpsta在转基因大豆中定量表达与疫霉根腐病和灰斑病抗性相关研究［J］.中国油料作物学报,2015,37(1):35-40.(Gu X N, Liu Z K,Wang P W, et al.Gene expression and disease resistance of transformed hrpZpsta gene in transgenic soybean［J］. Chinese Journal of Oil Crop Sciences, 2015, 37(1): 35-40.)［31］唐微.转基因拷贝数对农艺性状的影响［J］.安徽农业科学,2008,36(32):13991-13992.(Tang W. Effect of transgenic copy number on agronomic traits［J］. Journal of Anhui Agricultural Sciences,2008,36(32):13991-13992.)［32］Yang H, Singsit C, Wang A, et al. Transgenic peanut plants containing a nucleocapsid protein gene of tomato spotted wilt virus show divergent levels of gene expression［J］. Plant Cell Reports,1998, 17(9): 693-699.［33］华志华,朱雪峰,吴明国,等.水稻转基因整合模式中外源基因的遗传规律［J］.作物学报,2003,29(1):44-48.(Hua Z H,Zhu X F,Wu M G,et al.Inheritance of exo-genes integrated into the rice genomes［J］.Acta Agronomica Sinica, 2003, 29(1):44-48.)［34］刘娜,张锐,罗淑萍,等. 荧光定量PCR技术检测vgb基因在棉花中的表达［J］.新疆农业大学学报, 2007, 30(3): 6-9. (Liu N,Zhang R, Luo S P, et al. Expression level of detecting vgb gene by fluorescence real-time quantitative PCR in cotton［J］. Journal of Xinjiang Agricultural University, 2007, 30(3): 6-9.)

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Last Update: 2020-06-10