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当前位置: 首页 > 文章 > 一种快速鉴定柑橘黄龙病耐性种质材料的方法 南方农业学报 2019 (11) 2489-2495
Position: Home > Articles > A rapid identification method for germplasm materials resistant to citrus Huanglongbing disease Journal of Southern Agriculture 2019 (11) 2489-2495

一种快速鉴定柑橘黄龙病耐性种质材料的方法

作  者：
唐艳;武晓晓;娄兵海;刘萍;陈传武;牛英;张戈壁;燕佳文;邓崇岭
单  位：
广西师范大学生命科学学院;广西特色作物研究院/广西柑橘生物学重点实验室
关键词：
柑橘;黄龙病;耐性;种质材料;高接染毒鉴定法
摘  要：
【目的】研发一种快速鉴定柑橘黄龙病耐性种质材料的方法,为加快柑橘黄龙病耐性育种进程和提高育种效率打下基础。【方法】以收集的36份疑似对柑橘黄龙病具有耐性的柑橘种质为材料,采用直接高接于感染黄龙病菌柑橘树上的方法(高接染毒鉴定法),通过田间症状观察结合定量PCR检测对试验材料的耐病性进行鉴定评价。【结果】春季高接供试材料1个月后接芽萌发,3个月后(2018年6月5日)首次在KH-14上出现典型的叶片斑驳型黄化症状,4个月后(2018年7月5日)观察有23份种质材料的叶片出现斑驳型黄化症状,另外11份种质材料未表现症状;6个月后(2018年9月4日)观察发现KH-18、KH-12、KHY-4、KHY-5和KHY-6等5份种质材料的生长虽然较正常,但叶片已出现黄化症状,只有KH-21的枝梢生长良好,无黄梢和斑驳型黄化叶,初步判定为对黄龙病具有耐性的种质材料。对6份种质材料的定性PCR检测结果表明,KH-21为阴性,其他5份材料为阳性;实时荧光定量PCR检测结果,KH-21的平均黄龙病菌含量为1870.0个细胞/μg DNA,对照材料平均黄龙病菌含量为372285.5个细胞/μg DNA,表明KH-21对柑橘黄龙病具有耐性。【结论】高接染毒鉴定法鉴定周期在6个月左右,具有通量大、时间短、效率高、成本低及结果准确等优点,能达到快速鉴定柑橘品种对柑橘黄龙病耐性的目的,可在柑橘黄龙病耐性鉴定中推广使用。
译  名：
A rapid identification method for germplasm materials resistant to citrus Huanglongbing disease
作  者：
TANG Yan;WU Xiao-xiao;LOU Bing-hai;LIU Ping;CHEN Chuan-wu;NIU Ying;ZHANG Ge-bi;YAN Jia-wen;DENG Chong-ling;Guangxi Academy of Specialty Crops/Guangxi Key Laboratory of Citrus Biology;College of Life Science,Guangxi Normal University;
关键词：
citrus;;Huanglongbing disease;;resistance;;germplasm material;;method for infecting bacteria by top grafting
摘  要：
【Objective】To develop a rapid identification method for citrus Huanglongbing disease resistant germline materials and provide basis for accelarating the citrus breeding that were resistant to Huanglongbing and increasing the breeding efficiency.【Method】Thirty-six citrus germplasms suspected to be resistant to citrus Huanglongbing disease were collected in this study. The method of direct high grafting to citrus trees infected with Huanglongbing pathogen was adopted. The resistance of the test materials was identified and evaluated by field symptoms combined with quantitative PCR.It was defined as the top grafting identification method.【Result】The test materials that were grafted in spring started to germinate after one month,and three months later(June 5,2018),typical mottled yellowing on leaves was observed on KH-14 for the fiest time. After four months(July 5,2018)of top grafting,typical mottled yellowing occurred on 23 materials,and 11 materials showed no such symptom. After six months(September 4,2018)of top grafting,although the growth of KH-18,KH-12,KHY-4,KHY-5 and KHY-6 were normal,yellowing was observed on their leaves. Only KH-21 grew well,and showed no yellow shoots and yellowing leaves. It was identified as the material with resistance to Huanglongbing disease. Quantitative PCR tests on the above six materials showed that KH-21 was negative and the other five were positive. Real-time fluorescence quantitative PCR test indicated that the average Huanglongbing bacteria amount in KH-21 was 1870.0 cell/μg DNA,and the average Huanglongbing bacteria amount incontrol material was 372285.5 cell/μg DNA,indicating KH-21 was resistant to Huanglongbing bacteria.【Conclusion】The method for infecting bacteria by topgrafting takes six months,and can detect large amount of seedlings,it is time-saving,high efficient,low-cost and accurate. This methodcan quickly identify the resistance of citrus varieties to citrus Huanglongbing disease,and can be popularized and used in the identification of citrus Huanglongbing disease resistance.

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