甜瓜抗枯萎病资源筛选及抗病生理指标相关性分析

Screening of melon resources resistant to Fusarium wilt and correlation analysis of physiological indexes of disease resistance

摘要

为明确黑龙江地区甜瓜枯萎病生理小种，探究枯萎病胁迫下甜瓜生理指标变化，完善甜瓜枯萎病抗病评价体系，利用表型鉴定结合分子标记辅助选择(MAS，marker-assisted selection)对24份甜瓜资源进行抗病筛选。结果表明，黑龙江省尖孢镰刀菌甜瓜专化型Fm-1为1号生理小种。分子标记辅助选择共筛选出抗病资源15份，感病资源9份，具有较高的筛选效率。其中野生型资源1314抗性最强，发病率仅为10%；栽培种22-152的感病程度最高。利用筛选的两个抗性极端材料在幼苗期接种病原菌并于根部取样，测定抗病生理指标并进行相关性分析。结果表明，枯萎病胁迫下的抗病和感病材料生理指标响应差异显著，过氧化氢酶(CAT)、苯丙氨酸解氨酶(PAL)、超氧化物歧化酶(SOD)和过氧化物酶(POD)活性与发病率呈极显著负相关；丙二醛含量和相对电导率与发病率呈极显著正相关。综上，利用功能性分子标记可在早期实现精准筛选甜瓜抗枯萎病品种，相对电导率和CAT及PAL活性可作为抗枯萎病评价的首选指标。

Abstract

In order to identify the physiological race of melon Fusarium wilt in Heilongjiang province, and improve the evaluation system of melon Fusarium wilt disease resistance, 24 melon varieties were screened for disease resistance using phenotype identification combined with marker-assisted selection (MAS). The results showed that the specific type Fm-1 of Fusarium oxysporum from Heilongjiang province was physiological race No. 1. A total of 15 resistant resources and 9 susceptible resources were screened by marker-assisted selection with high screening efficiency. The wild-type resource 1314 had the most resistant, the incidence of which was only 10%. The cultivated-type 22-152 had the highest degree of infection. Two resources with extreme resistance were inoculated with pathogens at seedling stage and sampled at root. The physiological indexes of resistance were determined and the correlation analysis was carried out. The results showed that the physiological indexes of resistant and susceptible materials were significantly different under wilt stress, and the activities of catalase (CAT), phenylalanine aminolyase (PAL), superoxide dismutase (SOD) and peroxidase (POD) were significantly negatively correlated with incidence rate. Malondialdehyde content and relative conductivity were significantly positively correlated with incidence rate. In conclusion, functional molecular markers can be used to accurately screen melon varieties resistance to Fusarium wilt at an early stage, and relative conductivity and CAT and PAL activities can be used as the preferred indicators for evaluation of Fusarium wilt resistance.

关键词

甜瓜 / 枯萎病 / 抗病资源筛选 / 生理指标测定 / 相关性分析

Key words

Melon / Fusarium wilt / Screening of resistant resources / Physiological index determination / Correlation analysis

参考文献

[1] 栾非时. 西瓜甜瓜育种与生物技术[M]. 北京: 科学出版社, 2013.

[2] 王先挺, 曾立红, 王斌, 等. 大棚甜瓜枯萎病病原鉴定及其生物学特性[J]. 贵州农业科学, 2020, 48(7): 42-46.

[3] 刘春艳, 王万立, 郝永娟, 等. 大棚甜瓜枯萎病的发生及综合防治[J]. 农业科技通讯, 2010(7): 171-172.

[4] 张淑梅, 赵晓宇, 张先成, 等. 3种瓜类枯萎病菌 Fusarium oxysporum的分子检测[J]. 植物病理学报, 2010, 40(6): 636-641.

[5] 刘朋义, 别之龙, 彭斌, 等. 甜瓜品种抗枯萎病的苗期鉴定[J]. 中国瓜菜, 2011, 24(2): 11-13.

[6] SADEGHPOUR N, ASADI-GHARNEH H A, NASR-ESFAHANI M N, et al. Antioxidant enzymes associated with resistance to Fusarium oxysporum f. sp. melonis race 1.2 in melon[J]. Physiological and Molecular Plant Pathology, 2022, 121: 101880.

[7] 关峰, 万新建, 张景云, 等. 苦瓜枯萎病研究进展[J]. 中国瓜菜, 2018, 31(5): 1-4.

[8] RIOU C, FREYSSINET G, FEVRE M. Production of cell wall-degrading enzymes by the phytopathogenic fungus Sclerotinia sclerotiorum[J]. Applied and Environmental Microbiology, 1991, 57(5): 1478-1484.

[9] MUTHUKUMAR A, ESAWRAN A, SANGEETHA G. Induction of systemic resistance by mixtures of fungal and endophytic bacterial isolates against Pythium aphanidermatum[J]. Acta Physiologiae Plantarum, 2011, 33(5): 1933-1944.

[10] 赵秀娟, 唐鑫, 程蛟文, 等. 酶活性、丙二醛含量变化与苦瓜抗枯萎病的关系[J]. 华南农业大学学报, 2013, 34(3): 372-377.

[11] 邹芳斌, 司龙亭, 李新, 等. 黄瓜枯萎病抗性与防御系统几种酶活性关系的研究[J]. 华北农学报, 2008, 23(3): 181-184.

[12] 佘小漫, 何自福, 罗方芳, 等. 瓜类枯萎病及其综合防治技术[J]. 广东农业科学, 2011, 38(2): 84-87.

[13] 王彩霞, 陈晓林, 李保华. 腐烂病菌侵染对苹果愈伤组织防御酶活性及丙二醛含量的影响[J]. 植物生理学报, 2014, 50(7): 909-916.

[14] RISSER G, BANIHASHEMI Z, DAVIS D W. A proposed nomenclature of Fusarium oxysporum f. sp. melonis races and resistance genes in cucumis melo[J]. Phytopathology, 1976, 66(9): 1105-1106.

[15] HERMAN R, PERL-TREVES R. Characterization and inheritance of a new source of resistance to Fusarium oxysporum f. sp. melonis race 1.2 in cucumis melo[J]. Plant Disease, 2007, 91(9): 1180-1186.

[16] ZINK F W, GUBLER W D. Inheritance of resistance in muskmelon to Fusarium wilt[J]. Journal of the American Society for Horticultural Science, 1985, 110(5): 600-604.

[17] OUMOULOUD A, ARNEDO-ANDRES M S, GONZALEZ-TORRES R, et al. Inheritance of resistance to Fusarium oxysporum f. sp. melonis races 0 and 2 in melon accession tortuga[J]. Euphytica, 2010, 176(2): 183-189.

[18] TEZUKA T, WAKI K, KUZUYA M, et al. Development of new DNA markers linked to the Fusarium wilt resistance locus Fom-1 in melon[J]. Plant Breeding, 2011, 130(2): 261-267.

[19] JOOBEUR T, KING J J, NOLIN S J, et al. The Fusarium wilt resistance locus Fom-2 of melon contains a single resistance gene with complex features[J]. Plant Journal, 2004, 39(3): 283-297.

[20] SCHREUDER W, LAMPRECHT S C, HOLZ G. Race determination and vegetative compatibility grouping of Fusarium oxysporum f. sp. melonis from South Africa[J]. Plant Disease, 2000, 84(3): 231-234.

[21] MARTYN R D, BRUTON B D. An initial survey of the united states for races of Fusarium oxysporum f. sp. niveum[J]. HortScience, 1989, 24(4): 696-698.

[22] MARTYN R D. Fusarium oxysporum f. sp. niveum race 2: A highly aggressive race new to the United States[J]. Plant Disease, 1987, 71(3): 233-236.

[23] 凌悦铭, 李寐华, 杨永, 等. 基于BSA-Seq技术的甜瓜抗霜霉病InDel标记开发[J]. 新疆农业科学, 2021, 58(12): 2265-2273.

[24] 王士伟. 基于单核苷酸多态性的甜瓜枯萎病抗性基因 Fom-2功能性分子标记开发与利用[D]. 杭州: 浙江大学, 2010.

[25] 郝芳敏, 臧全宇, 丁伟红, 等. 浙江省甜瓜枯萎病生理小种鉴定及部分甜瓜品种抗性鉴定[J]. 浙江农业科学, 2023, 64(5): 1273-1276.

[26] 顾卫红, 王鸣. 甜瓜种质资源对枯萎病抗性的苗期人工接种鉴定[J]. 中国西瓜甜瓜, 1989(1): 1-8.

[27] 王怀松, 贺超兴, 张志斌, 等. 甜瓜白粉病抗性AFLP连锁标记的初步研究[J]. 中国瓜菜, 2009, 22(2): 4-6.

[28] ALPASLAN M, GUNES A. Interactive effects of boron and salinity stress on the growth, membrane permeability and mineral composition of tomato and cucumber plants[J]. Plant and Soil, 2001, 236(1): 123-128.

[29] 张翠梅, 师尚礼, 吴芳. 干旱胁迫对不同抗旱性苜蓿品种根系生长及生理特性影响[J]. 中国农业科学, 2018, 51(5): 868-882.

[30] PARIDA A K, DAS A B, MOHANTY P, et al. Defense potentials to NaCl in a mangrove, Bruguiera parviflora: Differential changes of isoforms of some antioxidative enzymes[J]. Journal of Plant Physiology, 2004, 161(5): 531-542.

[31] LIMON-PACHECO J, GONSEBATT M E. The role of antioxidants and antioxidant-related enzymes in protective responses to environmentally induced oxidative stress[J]. Mutation Research-genetic Toxicology and Environmental Mutagenesis, 2009, 674(1/2): 137-147.

[32] KUK Y I, SHIN J S, BURGOS N R, et al. Antioxidative enzymes offer protection from chilling damage in rice plants[J]. Crop Science, 2003, 43(6): 2109-2117.

[33] MANDAL S, MITRA A, MALLICK N. Biochemical characterization of oxidative burst during interaction between Solanum lycopersicum and Fusarium oxysporum f. sp. lycopersici[J]. Physiological and Molecular Plant Pathology, 2008, 72(1/3): 56-61.

[34] YOU J, CHAN Z L. ROS regulation during abiotic stress responses in crop plants[J]. Frontiers in Plant Science, 2015, 6: 1092.

[35] 丁玉梅, 张杰, 谢俊俊, 等. 枯萎病菌胁迫下3种黑籽南瓜 HQRGA2表达及抗氧化酶活性差异分析[J]. 植物生理学报, 2019, 55(3): 349-358.

[36] 刘佳, 徐秉良, 薛应钰, 等. 美洲南瓜(Cucurbita pepo)种皮苯丙氨酸解氨酶基因克隆与表达分析[J]. 中国农业科学, 2014, 47(6): 1216-1226.

[37] MADADKHAH E, LOTFI M, NABIPOUR A, et al. Enzymatic activities in roots of melon genotypes infected with Fusarium oxysporum f. sp. Melonis race 1[J]. Scientia Horticulturae, 2012, 135: 171-176.

[38] 邵元健, 周小林, 包卫红, 等. 甜瓜抗枯萎病基因 Fom-2分子标记及南通市枯萎病生理小种鉴定[J]. 扬州大学学报(农业与生命科学版), 2013(3): 90-94.

[39] 赵玉龙. 甜瓜橙色果肉、全雌系及主要抗病性状MAS的建立与评价[D]. 哈尔滨: 东北农业大学, 2017.

基金资助

黑龙江省农业科技创新跨越工程优青项目(CX22YQ32)

黑龙江省农业科技创新跨越工程农业特色产业项目(CX23TS11)

黑龙江省农业科学院院级课题(DQ-ZD-001)