小G蛋白StRab5b对马铃薯黄萎病的抗性功能

doi:10.7668/hbnxb.20195675

摘要/Abstract

摘要： 为了探究马铃薯小G蛋白StRab5b在黄萎病抗性中的作用,采用qRT-PCR法测定马铃薯组培苗(虎头、夏坡蒂)接种大丽轮枝菌后根系中StRab5b基因的相对表达量;以接种大丽轮枝菌后超表达StRab5b马铃薯株系L7、L8、L10为处理(StRab5b-L7、StRab5b-L8、StRab5b-L10),非转基因植株为对照(CK)检测其根系大丽轮枝菌的生物量、叶片病斑面积、过氧化氢(H₂O₂)含量、丙二醛(MDA)含量和抗氧化酶活性;以超表达StRab5b-L7株系和非转基因植株(CK)为材料利用盆栽法对其病情指数进行计算。结果表明,接种大丽轮枝菌后马铃薯诱导了StRab5b基因的表达,与接种0 h相比,接种后72 h,虎头和夏坡蒂的相对表达量分别显著提高701.12%,649.41%。接种大丽轮枝菌后,超表达StRab5b提高了马铃薯对黄萎病的抗性,与CK相比,接种后96 h,StRab5b-L7、StRab5b-L8、StRab5b-L10处理叶片病斑面积显著降低78.56%,66.56%,59.76%;接种后168 h,根系中大丽轮枝菌生物量分别显著降低73.07%,51.69%,23.38%;接种后96 h,MDA含量分别显著降低47.13%,33.08%,14.70%;接种后72 h,H₂O₂含量分别显著提高83.67%,48.38%,20.15%,CAT活性分别提高53.54%,33.28%,12.62%;接种后48 h,SOD活性分别显著提高53.01%,36.36%,16.54%;接种后96 h,POD活性分别提高55.58%,27.18%,1.73%;接种后72 h,APX活性分别显著提高63.69%,44.27%,17.11%。与CK相比,StRab5b-L7株系病株率和病情指数分别显著降低43.33百分点,86.07%。综上所述,超表达StRab5b基因后增强了马铃薯对黄萎病的抗性。

关键词: 马铃薯, 小G蛋白, 黄萎病, 抗病性

Abstract:

The aim was to explore the role of the small G protein StRab5b in potato resistance to Verticillium wilt.The relative expression levels of the StRab5b gene in the roots of potato tissue culture seedlings(Hutou and Xiapodi)after inoculation with Verticillium dahliae were determined by qRT-PCR.The biomass of V.dahliae in the roots,the lesion area on potato leaves,the content of hydrogen peroxide(H₂O₂),the content of malondialdehyde(MDA),and the activity of antioxidant enzymes were detected in the transgenic potato lines L7,L8,and L10 overexpressing StRab5b(StRab5b-L7,StRab5b-L8,StRab5b-L10)and the non-transgenic plants(CK)after inoculation with V.dahliae.The disease index was calculated using the pot culture method with the StRab5b-L7 transgenic line and the non-transgenic plants(CK).The results showed that the expression of the StRab5b gene was induced in potato after inoculation with V.dahliae.Compared with 0 h after inoculation,the relative expression levels of Hutou and Xiapodi increased significantly by 701.12% and 649.41% at 72 h after inoculation,respectively.After inoculation with V.dahliae,overexpression of StRab5b enhanced the resistance of potato to Verticillium wilt.Compared with CK,the lesion area on potato leaves in StRab5b-L7,StRab5b-L8,and StRab5b-L10 decreased significantly by 78.56%,66.56%,and 59.76% at 96 h after inoculation,respectively;the biomass of V.dahliae in the roots decreased significantly by 73.07%,51.69%,and 23.38% at 168 h after inoculation,respectively;the MDA content decreased significantly by 47.13%,33.08%,and 14.70% at 96 h after inoculation,respectively;the H₂O₂ content increased significantly by 83.67%,48.38%,and 20.15% at 72 h after inoculation,respectively,and the CAT activity increased by 53.54%,33.28%,and 12.62%,respectively;the SOD activity increased significantly by 53.01%,36.36%,and 16.54% at 48 h after inoculation,respectively;the POD activity increased by 55.58%,27.18%,and 1.73% at 96 h after inoculation,respectively;the APX activity increased significantly by 63.69%,44.27%,and 17.11% at 72 h after inoculation,respectively.Compared with CK,the disease rate and disease index of the StRab5b-L7 transgenic line decreased significantly by 43.33 percentage points and 86.07%,respectively.In conclusion,overexpression of the StRab5b gene enhanced the resistance of potato to Verticillium wilt.

Key words: Potato, Small G protein, Verticillium wilt, Disease resistance

中图分类号:

孙学桃, 伊海佼, 孙婧, 郑国华, 刘志达, 樊根, 赵君, 张之为. 小G蛋白StRab5b对马铃薯黄萎病的抗性功能[J]. 华北农学报, 2025, 40(6): 177-185. doi: 10.7668/hbnxb.20195675.

SUN Xuetao, YI Haijiao, SUN Jing, ZHENG Guohua, LIU Zhida, FAN Gen, ZHAO Jun, ZHANG Zhiwei. Resistance of Small G Protein StRab5b to Verticillium Wilt of Potato[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(6): 177-185. doi: 10.7668/hbnxb.20195675.

http://www.hbnxb.net/CN/Y2025/V40/I6/177

图/表 8

表1 qRT-PCR引物序列信息

Tab.1 qRT-PCR primer sequence information

引物名称 Primer name	引物序列(5'—3') Primer sequence(5'—3')
q-StActin-F	CACCCTGTTCTGCTCACT
q-StActin-R	CAGCCTGAATAGCAACATAC
Fg-qGFP-F	CGACAAGCAGAAGAACGGCATCAA
Fg-qGFP-R	CTTCTCGTTGGGGTCTTTGCTCA
q-StRab5b-F	ATAGCCTTGCAGGACTCAAC
q-StRab5b-R	CAGCACCTCGGTAGTATAATGG

图1 接种大丽轮枝菌后StRab5b基因的相对表达量不同小写字母表示差异显著(P<0.05)。图2—7同。

Fig.1 Relative expression of StRab5b gene after inoculation with Verticillium dahliae Different lowercase letters indicate significant differences(P<0.05).The same as Fig.2—7.

图2 接种后马铃薯叶片病斑面积变化 A.接种后马铃薯叶片病斑面积变化;B.接种后马铃薯叶片病斑面积定量分析。

Fig.2 Changes of lesion area on potato leaves after inoculation A.Changes of lesion area on potato leaves after inoculation;B.Quantitative analysis of potato leaves lesion area after inoculation.

图3 接种后马铃薯根系大丽轮枝菌的定殖情况 A.接种后马铃薯根系中的荧光强度;B.接种后马铃薯根系平均荧光强度定量分析;C.接种后马铃薯根系定殖的大丽轮枝菌生物量。

Fig.3 Colonization of Verticillium dahliae on potato roots after inoculation A.Fluorescence intensity in potato roots after inoculation;B.Quantitative analysis of average fluorescence intensity of potato root after inoculation;C.The biomass of Verticillium dahliae colonizing potato roots after inoculation.

图4 接种后马铃薯叶片MDA含量变化

Fig.4 Changes in MDA content in potato leaves after inoculation

图5 接种后马铃薯叶片过氧化氢含量变化 A.接种后马铃薯叶片DAB染色;B.接种后马铃薯叶片过氧化氢含量变化。

Fig.5 Changes in hydrogen peroxide content in potato leaves after inoculation A.DAB staining of potato leaves after inoculation;B.Changes in hydrogen peroxide content in potato leaves after inoculation.

图6 接种后马铃薯叶片CAT、SOD、POD、APX活性

Fig.6 Activities of CAT,SOD,POD and APX in potato leaves after inoculation

图7 盆栽苗接种大丽轮枝菌后发病情况 A.株系发病症状。

Fig.7 Incidence of potted plants inoculated with Verticillium dahlia A.Phenotypic symptoms of the strain.

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