新疆北疆部分棉区落叶型棉花黄萎病菌落形态特征及致病力鉴定

doi:10.6048/j.issn.1001-4330.2025.01.020

新疆农业科学 ›› 2025, Vol. 62 ›› Issue (1): 174-181.DOI: 10.6048/j.issn.1001-4330.2025.01.020

新疆北疆部分棉区落叶型棉花黄萎病菌落形态特征及致病力鉴定

赖成霞¹(), 杨延龙¹, 汪鹏龙¹, 朱梦宇², 杨栋³, 李春平¹, 葛风伟⁴, 玛依拉·玉素音¹, 阳妮¹, 马君¹()

1.新疆农业科学院经济作物研究所,乌鲁木齐 830091
2.新疆农业大学农学院,乌鲁木齐 830091
3.新疆维吾尔自治区植物保护站,乌鲁木齐 830049
4.新疆师范大学生命科学学院,乌鲁木齐 830054

收稿日期:2024-07-27 出版日期:2025-01-20 发布日期:2025-03-11
通信作者: 马君(1983-),男,甘肃民勤人,副研究员,研究方向为棉花种质资源收集与利用,(E-mail)xj.majun@163.com
作者简介:赖成霞(1972-),女,新疆乌鲁木齐人,副研究员,研究方向为棉花抗逆分子育种,(E-mail)lchxia2001@163.com
基金资助:
新疆维吾尔自治区重大专项“新疆主要农作物创新育种工程-子课题机采棉新品种培育及技术示范”(2021A02001-4)

Pathogen colony’s morphological characteristics and pathogenicity identification of defioating cotton Verticillium wilt in some areas in northern Xinjiang

LAI Chengxia¹(), YANG Yanlong¹, WANG Penglong¹, ZHU Mengyu², YANG Dong³, LI Chunping¹, GE Fengwei⁴, Mayila Yusuyin¹, YANG Ni¹, MA Jun¹()

1. Research Institute of Economic Crops, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China
2. College of Agriculture, Xinjiang Agricultural University, Urumqi 830091, China
3. Plant Protection Station of Xinjiang Uygur Autonomous Region, Urumqi 830049, China
4. College of Life Sciences, Xinjiang Normal University, Urumqi 830054, China

Received:2024-07-27 Published:2025-01-20 Online:2025-03-11
Supported by:
Major Scientific R & D Program Project of Xinjiang Uygur Autonomous Region“Cultivation and Technology Demonstration of New Varieties of Machine-Picked Cotton”(2021A02001-4)

摘要/Abstract

摘要：

【目的】 分析新疆北疆地区棉花黄萎病的致病力及其分化现状,以昌吉市、沙湾市为主,鉴定两地收集的棉花黄萎病的形态特征及致病力。【方法】 利用棉花落叶型黄萎病的特异性引物DB19f/DB22r和非落叶型引物INTND2f/INTND2r对所有菌株的致病力进行分子鉴定,并进行全基因组重测序,以SNP 位点作为标记进行遗传多样性分析,并利用显微镜观察菌落、菌丝和孢子形态,以强致病力菌种V991作对照,观察菌落形态和致病力鉴定,采用叶绿素荧光成像技术比较观测具有强致病力的Vd-3和V991的病情变化图像和荧光参数变化趋势。【结果】 14个菌株均为落叶型。其菌落为圆形,菌丝的轮枝数为2,孢子形态为长卵圆形。与V991相比,Vd-3的病情指数最高,属强致病力的菌株,其对棉花具有明显不同的侵染方式和速度,Vd-3的侵染从叶缘到叶片中央,而V991的侵染是从叶肉部分的零星发病扩展到叶缘,再发展到中央部分,并且Vd-3侵染叶片各荧光成像和参数天变化趋势更为显著,其中非调节性能量耗散的量子产量Y(NO)和非光化学淬灭NPQ4在第6 d即到变化拐点,最大光化学量子产量Fv/Fm和实际光量子产量Y(Ⅱ)则变化缓慢,13 d 时Fv/Fm表现明显差异。【结论】 昌吉市、沙湾市两地的大丽轮枝菌均为落叶型,圆形菌落,菌丝轮枝数是2,长卵圆形孢子,Vd-3不仅为遗传距离最远且变异性大的菌株,而且为最强致病力的黄萎病病株,与V991相比,对棉花具有不同的侵染方式,荧光参数Y(NO)和NPQ变化更为敏锐。

关键词: 致病力; 叶绿荧光成像; 棉花黄萎病; 全基因组重测序

Abstract:

【Objective】 To identify morphological characteristics and pathogenicity of cotton Verticillium wilt collected from Changji and Shawan in the hope of comprehending the pathogenicity and differentiation of the cotton disease in northern Xinjiang. 【Methods】 The molecular detections of the pathogenicity for cotton wilt disease were performed using specific primers DB19f/DB22r for defioating pathotype and INTND2f/INTND2r for nondefioating pathotype. Whole genome resequencing was performed using SNP loci as markers for genetic diversity analysis. The colony morphology and the pathogenicity were identified adopting the virulence strain V991 as the control, chlorophyll fluorescence imaging technology was employed to compare and observe the disease changes and the trend of fluorescence parameters of Vd-3 and V991. 【Results】 Molecular identification showed that all 14 strains examined were defioating pathotype. In comparison with V991 strain, Vd-3 exhibited the highest disease index and high pathogenicity with different infection modes and rapid infection rates; The infection of Vd-3 was seen that the disease symptom extended from the leaf margin to the center, while the that of V991 spread from the sporadic distribution of the mesophyll to the leaf margin and then to the central part by observing the dynamic changes of imaging and fluorescence parameters employing the chlorophyll fluorescence imaging system. The daily variation trend of fluorescence imaging and parameters infected by Vd-3 plants was more obvious; The quantum yield Y (NO) of non-regulatory energy dissipation and the non-photochemical quenching (NPQ) reached the inflection point on the 6th day. The maximum photochemical quantum yield (Fv/Fm) and the actual optical quantum yield Y (Ⅱ) changed slowly, and the Fv/Fm showed obvious difference at 13 d. 【Conclusion】 The strains from Changji and Shawan city are defioating pathotype, and the colony is circular with 2 hyphal branches and long oval spores. In addition to the greatest genetic distance and variability, the most virulent strain of Verticillium wilt disease is Vd-3. Compared with that of V991,Vd-3 possesses the varied the infection mode for cotton, and fluorescence parameters Y (NO) and NPQ can exquisitely perceive the changes.

Key words: pathogenicity; chlorophyll fluorescence imaging; cotton Verticillium wilt; whole genome resequencing

中图分类号:

S435.1

赖成霞, 杨延龙, 汪鹏龙, 朱梦宇, 杨栋, 李春平, 葛风伟, 玛依拉·玉素音, 阳妮, 马君. 新疆北疆部分棉区落叶型棉花黄萎病菌落形态特征及致病力鉴定[J]. 新疆农业科学, 2025, 62(1): 174-181.

LAI Chengxia, YANG Yanlong, WANG Penglong, ZHU Mengyu, YANG Dong, LI Chunping, GE Fengwei, Mayila Yusuyin, YANG Ni, MA Jun. Pathogen colony’s morphological characteristics and pathogenicity identification of defioating cotton Verticillium wilt in some areas in northern Xinjiang[J]. Xinjiang Agricultural Sciences, 2025, 62(1): 174-181.

图/表 7

图1 棉花黄萎病致病力分子鉴定电泳注:M:DNA Mark, 1:落叶型引物对DB19f/DB22r,2:非落叶型引物对INTND2f/INTND2r

Fig.1 Electrophoretic map of the molecular identification for cotton Verticillium wilt Notes:M:DNA Mark 1 defoliating primer pairs DB19f/DB22r, 2: non-defoliating primer pairsINTND2f/INTND2r

图2 14 份棉花黄萎病原菌的 N-J 进化树

Fig.2 The neighbor-joining phylogenetic tree for the strains of cottonVerticillium wilt

表1 不同菌落形态与显微结构

Tab.1 The morphology and microstructure of different colony

序号 ID	名称 Name	轮枝数 Number of round branches	孢子颜色 Conidia color	孢子形状 Conidia shape	孢子大小 Conidiasize (μm)	菌落颜色 Colony color	菌落形状 Colony shape
1	Vc-1	2	无色	卵圆形	(1.81-5.22)×(4.29-11.95)	白色	圆
2	Vc-2	2	无色	卵圆形	(2.05-6.36)×(4.42-11.15)	白色	圆
3	Vc-3	2	无色	卵圆形	(1.76-5.63)×(4.48-8.97)	白色	圆
4	Vc-4	2	无色	卵圆形	(2.25-5.79)×(3.56-9.32)	白色	圆
5	Vd-1	2	无色	卵圆形	(1.27-5.24)×(3.26-10.34)	白色	圆
6	Vd-2	2	无色	卵圆形	(2.48-5.12)×(4.23-9.46)	白色	圆
7	Vd-3	2	无色	卵圆形	(2.36-5.47)×(4.35-9.49)	黑色	圆
8	Vd-4	2	无色	卵圆形	(2-5.51)×(4.73-9.1)	白色	圆
9	Vd-5	2	无色	卵圆形	(2.29-8.51)×(1.45-9.66)	白色	圆
10	Vd-6	2	无色	卵圆形	(1.94-5.71)×(3.89-9.76)	白色	圆
11	Vd-7	2	无色	卵圆形	(1.94-6.5)×(5.36-9.93)	黑色	圆
12	Vd-8	2	无色	卵圆形	(1.76-4.39)×(4.57-9.44)	黑色	圆
13	Vd-9	2	无色	卵圆形	(2.29-5.59)×(4.46-10.34)	黑色	圆
14	Vd-10	2	无色	卵圆形	(1.59-5.49)×(4.48-9.19)	黑色	圆
15	V991	2	无色	卵圆形	(2-6.25)×(5.03-10.45)	白色	圆

图3 各菌落培养特性及显微观察

Fig.3 Culture characteristics and microscopic observation of each colony

图4 棉花叶片接种各菌系后10 d的病情指数

Fig.4 The disease index of cotton leaves inoculated by each Verticillium strain after 10 days

图5 Vd-3和V991侵染棉花叶片的外观特征及叶绿素荧光成像

Fig.5 Appearance characteristics and chlorophyll fluorescence imaging of cotton leaves infected with Vd-3 and V991

图6 接种大丽轮枝菌Vd-3和V991后的叶绿素荧光参数动力学曲线变化

Fig.6 Dynamics curve changes of chlorophyll fluorescence parameters after inoculation with Verticillium dahliae Vd-3 and V991

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新疆北疆部分棉区落叶型棉花黄萎病菌落形态特征及致病力鉴定

Pathogen colony’s morphological characteristics and pathogenicity identification of defioating cotton Verticillium wilt in some areas in northern Xinjiang

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