[1]陈曦,冯辉,张金凤,等.拟南芥抗根结线虫和蚜虫的功能基因鉴定[J].江苏农业学报,2015,(06):1250-1256.[doi:doi:10.3969/j.issn.1000-4440.2015.06.009]
 CHEN Xi,FENG Hui,ZHANG Jin-feng,et al.Identification of genes resistant to root knot nematodes and aphids in Arabidopsis thaliana[J].,2015,(06):1250-1256.[doi:doi:10.3969/j.issn.1000-4440.2015.06.009]
点击复制

拟南芥抗根结线虫和蚜虫的功能基因鉴定()
分享到:

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年06期
页码:
1250-1256
栏目:
植物保护
出版日期:
2015-12-31

文章信息/Info

Title:
Identification of genes resistant to root knot nematodes and aphids in Arabidopsis thaliana
作者:
陈曦冯辉张金凤卢小雪陈怀谷魏利辉
(江苏省农业科学院植物保护研究所,江苏南京210014)
Author(s):
CHEN XiFENG HuiZHANG Jin-fengLU Xiao-xueCHEN Huai-guWEI Li-hui
(Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China)
关键词:
根结线虫蚜虫拟南芥激活标签突变体抗性鉴定
Keywords:
root knot nematodeaphidArabidopsis activation tag mutantresistance identification
分类号:
S433.3,S433.89
DOI:
doi:10.3969/j.issn.1000-4440.2015.06.009
文献标志码:
A
摘要:
为了鉴定出增强植物对根结线虫和蚜虫抗性的功能基因,从9个抗蚜虫的拟南芥激活标签突变体中筛选增强根结线虫抗性的突变体。利用反向PCR技术定位激活标签在基因组上的插入位点,并通过荧光定量PCR技术分析插入位点上、下游基因表达水平,结合根结线虫和蚜虫在过量表达转基因植株上的表现验证基因功能。结果显示接种根结线虫后7 d,侵入3个拟南芥突变体根系的根结线虫数量明显少于野生型。其中2个突变体中的激活标签插入位点只相差2个碱基,SKS13基因表达水平提高;另一个株系中激活标签导致PERK2基因的表达水平提高。与野生型植株相比,分别过量表达SKS13和PERK2的转基因植株能明显减少侵入根系的根结线虫数量,同时蚜虫的繁殖数量也较低。
Abstract:
To identify plant genes that can increase resistance towards root knot nematode (RKN) and aphid, nine Arabidopsis activation tag mutants carrying aphid resistance were used to explore RKN resistance. Inverse PCR was performed to locate the activation tag on Arabidopsis genome and quantitative RT-PCR was used to quantify the expression levels of upstream and downstream genes. Three out of the nine aphid resistant mutants were identified with resistance towards RKN, indicated by lower infection level compared to wild type at 7 days after inoculation. Among these three mutants, two had almost the same location of the activation tag and activated SKS13 gene. In the other mutant, activation tag enhanced the expression level of PERK2 gene. Transgenic lines overexpressing either SKS13 or PERK2 resulted in reduced infection level of RKN and poorer aphid performance.

参考文献/References:

[1]万景旺,邵颖,朱华,等. 生防菌Jdm2 与生物源农药混用防治黄瓜根结线虫病的效果[J]. 江苏农业科学,2014,42(4): 108-110.
[2]高玉红,闫生辉,赵卫星. 印楝素与不同杀虫剂混配对根结线虫的防治效果[J]. 江苏农业科学,2014,42(7): 133-135.
[3]NICOL J M, TURNER S J, COYNE D L, et al., Current nematode threats to world agriculture, in genomics and molecular genetics of plant-nematode interactions[M]. Netherlands:Springer, 2011:21-43.
[4]KALOSHIAN I, DE ILARDUYA O M. Mi-1, a dual function disease resistance gene in tomato[C]// Keen N T, Mayama S. Delivery and Perception of Pathogen Signals in Plants. St Paul, MN:APS Press, 2001: 145-153.
[5]尤江,郭宏霞,张玉粉,等. 野生与人工种植伏毛铁棒锤对枸杞蚜虫的杀虫活性比较[J]. 江苏农业科学,2013,41(8):148-149.
[6]YAGHOOBI J, KALOSHIAN I, WEN Y, et al. Mapping a new nematode resistance locus in Lycopersicon peruvianum[J].Theoretical and Applied Genetics, 1995, 91(3): 457-464.
[7]KALOSHIAN I, YAGHOOBI J, LIHARSKA T, et al. Genetic and physical localization of the root-knot nematode resistance locus Mi in tomato[J]. Molecular and General Genetics MGG, 1998, 257(3): 376-385.
[8]MILLIGAN S B, BODEAU J, YAGHOOBI J, et al. The root knot nematode resistance gene Mi from tomato is a member of the leucine zipper, nucleotide binding, leucine-rich repeat family of plant genes[J].The Plant Cell, 1998, 10(8): 1307-1319.
[9]ROSSI M, GOGGIN F L, MILLIGAN S B, et al. The nematode resistance gene Mi of tomato confers resistance against the potato aphid[J]. Proceedings of the National Academy of Sciences, 1998, 95(17): 9750-9754.
[10]DE ILARDUYA O M, XIE Q G, KALOSHIAN I. Aphid-induced defense responses in Mi-1-mediated compatible and incompatible tomato interactions[J]. Molecular Plant-Microbe Interactions, 2003, 16(8): 699-708.
[11]DE ILARDUYA O M, MOORE A E, KALOSHIAN I. The tomato Rme1 locus is required for Mi-1-mediated resistance to root-knot nematodes and the potato aphid[J]. The Plant Journal, 2001, 27(5): 417-425.
[12]BHATTARAI K K, LI Q, LIU Y L, et al. The Mi-1-mediated pest resistance requires Hsp90 and Sgt1[J]. Plant Physiology, 2007, 144(1): 312-323.
[13]BHATTARAI K K, ATAMIAN H S, KALOSHIAN I, et al. WRKY72-type transcription factors contribute to basal immunity in tomato and Arabidopsis as well as gene-for-gene resistance mediated by the tomato R gene Mi-1[J]. The Plant Journal, 2010, 63(2): 229-240.
[14]ATAMIAN H, EULGEM T, KALOSHIAN I. SlWRKY70 is required for Mi-1-mediated resistance to aphids and nematodes in tomato[J].Planta, 2012, 235(2): 299-309.
[15]MANTELIN S, PENG H C, LI B B, et al. The receptor-like kinase SlSERK1 is required for Mi-1-mediated resistance to potato aphids in tomato[J].The Plant Journal, 2011, 67(3): 459-471.
[16]JABLONSKA B, AMMIRAJU J S S, BHATTARAI K K, et al. The Mi-9 gene from solanum arcanum conferring heat-stable resistance to root-knot nematodes is a homolog of Mi-1[J]. Plant Physiology, 2007, 143(2): 1044-1054.
[17]MARSCH-MARTINEZ N, GRECO R, VAN ARKEL G, et al. Activation tagging using the En-I maize transposon system in Arabidopsis[J]. Plant Physiology, 2002, 129(4): 1544-1556.
[18]CHEN X, VOSMAN B, VISSER R G, et al. High throughput phenotyping for aphid resistance in large plant collections[J]. Plant Methods, 2012, 8(1): 33-33.
[19]THIES J A, MERRILL S B, CORLEY E L. Red food coloring stain: new, safer procedures for staining nematodes in roots and egg masses on root surfaces[J]. Journal of Nematology, 2002, 34(2): 179-181.
[20]SHIMADA T L, SHIMADA T, HARA-NISHIMURA I. A rapid and non-destructive screenable marker, FAST, for identifying transformed seeds of Arabidopsis thaliana[J]. The Plant Journal, 2010, 61(3): 519-528.
[21]CHEN X, ZHANG Z, VISSER R, et al. Constitutive overexpression of the pollen specific gene SKS13 in leaves reduces aphid performance on Arabidopsis thaliana[J]. BMC Plant Biology, 2014, 14(1): 1-10.
[22]CHITWOOD D J. Research on plant-parasitic nematode biology conducted by the United States Department of Agriculture-Agricultural Research Service[J]. Pest Management Science, 2003, 59(6-7): 748-753.
[23]TJALLINGII W F. Continuous recording of stylet penetration activities by aphids, in Aphid-plant genotype interactions[M]. Amsterdam: Elsevier Science, 1990:89-99.
[24]LALOI C, APEL K, DANON A. Reactive oxygen signalling: the latest news[J]. Current Opinion in Plant Biology, 2004, 7(3): 323-328,
[25]ELLIS C, TURNER J G. The Arabidopsis mutant cev1 has constitutively active jasmonate and ethylene signal pathways and enhanced resistance to pathogens[J]. The Plant Cell, 2001, 13(5): 1025-1033.
[26]BHATTARAI K K, XIE Q G, MANTELIN S, et al. Tomato susceptibility to root-knot nematodes requires an intact jasmonic acid signaling pathway[J]. Molecular Plant-Microbe Interactions, 2008, 21(9): 1205-1214.
[27]BAI L, ZHANG G Z, ZHOU Y, et al. Plasma membrane-associated proline-rich extensin-like receptor kinase 4, a novel regulator of Ca2+ signalling, is required for abscisic acid responses in Arabidopsis thaliana[J]. The Plant Journal, 2009, 60(2): 314-327.
[28]SILVA N, GORING D. The proline-rich, extensin-like receptor kinase-1 (PERK1) gene is rapidly induced by wounding[J]. Plant Molecular Biology, 2002, 50(4-5): 667-685.
[29]LAMESCH P, BERARDINI T Z, LI D H, et al. The Arabidopsis information resource (TAIR): improved gene annotation and new tools[J]. Nucleic Acids Research, 2012,40(1): 1202-1210.

相似文献/References:

[1]李亚茹,王银磊,赵丽萍,等.番茄ty-5和 Mi-1基因多重 PCR 体系的建立[J].江苏农业学报,2016,(04):869.[doi:10.3969/j.issn.100-4440.2016.04.024]
 LI Ya-ru,WANG Yin-lei,ZHAO Li-ping,et al.Identification of ty-5 gene and Mi-1 gene by multiplex PCR in tomato[J].,2016,(06):869.[doi:10.3969/j.issn.100-4440.2016.04.024]
[2]范亚磊,赵敏,邓晟,等.侵染江苏猕猴桃的北方根结线虫(Meloidogyne hapla)形态学描述和分子特征分析[J].江苏农业学报,2021,(01):75.[doi:doi:10.3969/j.issn.1000-4440.2021.01.010]
 FAN Ya-lei,ZHAO Min,DENG Sheng,et al.Morphological description and molecular characteristic analysis on Meloidogyne hapla which infect kiwifruit in Jiangsu province[J].,2021,(06):75.[doi:doi:10.3969/j.issn.1000-4440.2021.01.010]
[3]李东育,韩大勇,徐养诚,等.不同人工干预方式下苹果园蚜虫-天敌群落结构及发生动态[J].江苏农业学报,2021,(04):848.[doi:doi:10.3969/j.issn.1000-4440.2021.04.005]
 LI Dong-yu,HAN Da-yong,XU Yang-cheng,et al.Community structure and occurrence dynamics of aphids and natural enemies in apple orchard under different artificial interventions[J].,2021,(06):848.[doi:doi:10.3969/j.issn.1000-4440.2021.04.005]

备注/Memo

备注/Memo:
收稿日期:2015-06-24 基金项目:公益性行业(农业)科研专项基金项目(201403032);2014江苏省博士计划项目(2014) 作者简介:陈曦(1983-),女,江苏滨海人,博士,助理研究员,主要从事植物抗病性研究。(Tel)025-4392995;(E-mail)chenxi_201402@163.com
更新日期/Last Update: 2015-12-31