黄瓜抗白粉病突变体筛选与鉴定

doi:10.3864/j.issn.0578-1752.2020.01.016

摘要/Abstract

摘要：

【目的】白粉病是危害黄瓜产量、品质的最严重的病害之一,抗白粉病材料的发掘与研究,可以实现从根本上解决病害问题。对获得的‘长春密刺’突变体材料进行分析,旨在筛选出黄瓜抗白粉病突变体材料,丰富育种群体。【方法】对400份‘长春密刺’突变体材料进行苗期接种白粉病菌试验,通过叶片病斑观察结合病情指数分析,初步筛选出抗病材料,将筛选出的抗病材料在大田环境下自然发病进一步观察表型。对初步筛选出的抗病材料进行苗期生理鉴定,测定并分析超氧化物歧化酶、过氧化物酶、过氧化氢酶活性以及叶绿素a、叶绿素b、类胡萝卜素、可溶性蛋白含量等生理指标,通过田间自然发病进一步筛选抗病材料,并测定其蒸腾速率、胞间二氧化碳浓度、气孔导度、净光合速率等光合作用指标及乙烯、茉莉酸、水杨酸激素含量,并通过荧光定量PCR分析叶片中乙烯、茉莉酸、水杨酸、木质素、病程相关蛋白等防御信号途径中相关基因的表达。【结果】与感病材料相比,抗病材料的表面菌斑少,净光合速率、气孔导度都优于对照‘长春密刺’,胞间CO₂浓度低于‘长春密刺’。在防御激素方面,抗病材料的乙烯、茉莉酸、水杨酸含量同样优于感病材料,而在成熟期叶片的防御信号途径相关基因表达上,抗病材料的表达高于感病材料。通过苗期接种白粉病菌和田间自然发病综合筛选出了两份抗白粉病突变体材料：Mu-86-2、Mu-58-9。【结论】通过对突变体库的筛选可以获得抗白粉病新材料,这些材料的获得对黄瓜抗白粉病的遗传研究和新品种培育具有重要价值。

关键词: 黄瓜, 突变体, 抗白粉病, 筛选鉴定

Abstract:

【Objective】Powdery mildew is one of the most serious diseases that effects cucumber yield and quality. The discovery and research of materials resistant to powdery mildew can fundamentally solve the disease problem. To screen out materials of cucumber resistant to powdery mildew and enrich the breeding population, the Changchunmici mutants were investigated and analyzed in the study.【Method】Totally, 400 Changchunmici mutant lines were inoculated with powdery mildew at seedling stage, and resistant materials were screened preliminarily based on leaf lesion observation combined with disease index analysis. The selected resistant materials were further observed in the natural environment in the field environment. Physiological indexes of resistant materials in the seedling stages were analyzed, including the activities of superoxide dismutase, peroxidase and catalase activity, and physiological indicators such as the contents of chlorophyll a, chlorophyll b, carotenoids and soluble protein. The resistant materials were screened by natural pathogenesis in the field and were further analyzed for the photosynthetic indexes such as transpiration rate, intercellular carbon dioxide concentration, stomatal conductance and net photosynthetic rate. And the contents of ethylene, jasmonic acid and salicylate were determined, and expression of related genes in defense signaling pathways such as ethylene, jasmonic acid, salicylic acid, lignin, and disease-related proteins in leaves were analyzed by real-time PCR. 【Result】Compared with susceptible materials, the resistant materials had less plaque. The net photosynthetic rate and stomatal conductance of resistant materials were higher than those of wild-type Changchunmici, and the intercellular CO₂ concentration of them was lower than that of Changchunmici. In terms of defensive hormones, the contents of ethylene, jasmonic acid and salicylate of resistant materials were higher than those in control. The expression of defense signal related genes in mature leaves of resistant materials was higher compared with susceptible materials. Two powdery mildew resistance materials, Mu-86-2 and Mu-58-9 were screened by inoculation of powdery mildew in seedling stage and natural disease. 【Conclusion】The new materials resistant to powdery mildew could be obtained by screening mutant libraries. The acquisition of these materials had the important value for the genetic research and new varieties breeding of cucumber resistance to powdery mildew.

Key words: cucumber, mutant, resistance to powdery mildew, screening and identification

亓飞,林姝,宋蒙飞,张孟茹,陈姝延,张乃心,陈劲枫,娄群峰. 黄瓜抗白粉病突变体筛选与鉴定[J]. 中国农业科学, 2020, 53(1): 172-182.

Fei QI,Shu LIN,MengFei SONG,MengRu ZHANG,ShuYan CHEN,NaiXin ZHANG,JinFeng CHEN,QunFeng LOU. Screening and Identification of Cucumber Mutant Resistant to Powdery Mildew[J]. Scientia Agricultura Sinica, 2020, 53(1): 172-182.

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编号Code	病情指数Disease index	抗性类型Disease resistance	编号Code	病情指数Disease index	抗性类型Disease resistance
Mu-86-2	2.22	抗R	Mu-104-2	20.20	中抗MR
Mu-58-9	3.33	抗R	Mu-106-3	22.22	中抗MR
Mu-50-10	4.18	抗R	Mu-59-8	22.22	中抗MR
Mu-9-9	9.00	中抗MR	Mu-150-3	23.23	中抗MR
Mu-60-5	10.00	中抗MR	Mu-118-2	24.24	中抗MR
Mu-84-8	15.56	中抗MR

编号 Code	叶绿素a含量 Chlorophyll a content	叶绿素b含量 Chlorophyll b content	类胡萝卜素 Carotenoids content
CK	13.556±1.089abcd	4.713±1.081b	2.420±0.987b
Mu-86-2	14.869±0.521abc	5.536±0.179ab	2.946±0.195ab
Mu-58-9	16.723± 0.536a	6.298±0.504ab	2.887±0.484ab
Mu-50-10	11.825±3.908d	4.566±0.756ab	2.501±0.942b
Mu-9-9	11.597±2.778bcd	4.291±0.886b	2.526±0.450b
Mu-60-5	14.317±2.577abc	5.342±0.709ab	2.853±0.356ab
Mu-84-8	14.921±2.864abc	5.635±1.691ab	2.950±0.296ab
Mu-104-2	11.845±1.490cd	4.557±0.526b	2.609±0.475b
Mu-106-3	14.333±2.758abc	5.599±0.923ab	2.993±0.292ab
Mu-59-8	15.372±3.327abc	5.930±1.075ab	3.042±0.938a
Mu-150-3	14.749±0.876abc	5.594±0.886ab	2.858±0.466ab
Mu-118-2	15.839±1.838ab	6.100±0.822a	2.783±0.185ab

编号Code	POD活性POD activity (U·g^-1·min^-1)	SOD活性SOD activity (U·g^-1)	CAT活性CAT activity (U·g^-1·min^-1)
CK	0.876±0.011bc	0.331±0.041bcde	13.104±0.381c
Mu-86-2	2.143±0.651a	0.450±0.108b	16.906±0.565ab
Mu-58-9	2.186±0.311a	0.415±0.028bc	18.800±2.630a
Mu-50-10	1.836±0.526b	0.588±0.024a	14.960±0.876bc
Mu-9-9	1.588±0.503b	0.309±0.003cde	13.120±0.155c
Mu-60-5	1.208±0.520b	0.358±0.085bcde	14.040±0.501bc
Mu-84-8	1.245±0.497b	0.302±0.115bcde	18.506±0.966a
Mu-104-2	0.832±0.119bc	0.327±0.049bcde	11.920±0.969d
Mu-106-3	0.589±0.237c	0.441±0.091bc	15.093±1.927ab
Mu-59-8	2.272±0.404a	0.247±0.046e	13.040±0.855c
Mu-150-3	0.977±0.228bc	0.268±0.087de	14.640±1.222bc
Mu-118-2	0.964±0.152bc	0.380±0.031bcd	14.340±0.650bc

编号 Code	净光合速率 Net photosynthetic rate (μmol·m^-2·s^-1)	气孔导度 Stomatal conductance (mmol·m^-2·s^-1)	胞间CO₂浓度 Intercellular CO₂ concentration (μmol·mol^-1)	蒸腾速率 Transpiration rate (mmol·m^-2·s^-1)
CK	16.500±1.252b	0.569±0.042b	302.666±9.865a	10.866±0.351a
Mu-58-9	22.866±1.625a	1.173±0.188a	286.000±1.000b	10.666±0.650a
Mu-86-2	22.866±2.281a	0.824±0.155b	278.000±10.000b	10.123±0.849a
IL52	23.600±0.435a	0.789±0.074 b	295.666±3.780b	11.133±0.416a

编号 Code	茉莉酸含量 Jasmonic acid content (pmol·L^-1)	水杨酸含量 Salicylic acid content (pmol·L^-1)	乙烯含量 Ethylene content (ng·L^-1)
CK	1346.912±25.758b	960.066±23.283d	159.509±3.868d
Mu-58-9	1696.515±31.178a	1114.259±19.026b	189.744±5.398b
Mu-86-2	1752.818±51.115a	1063.894±29.433a	194.694±6.823b
IL52	1692.587±47.625a	1009.656±10.652c	222.705±5.158a