甘肃马铃薯晚疫病菌群体结构及病害治理技术研究
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摘要
马铃薯是甘肃农业发展的战略性主导产业。由致病疫霉Phytophthorainfestans de Bary引起的马铃薯晚疫病近年来在马铃薯主产区频频发生，造成严重损失。致病疫霉群体表型结构组成、品种抗病性、环境因子决定了病害的流行程度。本研究以甘肃省马铃薯晚疫病为主要研究对象，从致病疫霉表型结构、土壤越冬卵孢子的活力与萌发、药剂的田间选择、品种抗病性测定和病害流行预警指导田间防控几方面进行研究，制定马铃薯晚疫病综合治理技术。
1.采用国际已知含单显性抗病基因系列品系R1-R11对采自甘肃马铃薯不同生态种植区的75个菌株进行了生理小种的鉴定，共鉴定出45个生理小种,其中生理小种l.2.3.4.5.6.7.8.9.l0.11出现频率最高，占被测菌株总数的22.67％，是甘肃省的优势生理小种。
2．对186个菌株进行甲霜灵敏感性测定，28.5％对甲霜灵表现敏感、15.6％表现中抗和55.9％表现高抗。各采集点均发现对甲霜灵表现高抗的菌株，这表明甘肃省马铃薯晚疫病菌对甲霜灵产生抗药性的频率比较高。
3.利用已知的A1、A2交配型标准菌株，对147个晚疫病菌株的交配型进行了测定，发现甘肃省马铃薯晚疫病菌交配型有A1、A2、A1A2、SF（自育型）4类，分别占被测菌株的66.2%、20.4%、2.1%、16.3%。
4.病原菌卵孢子存活和萌发试验发现在A1和A2交配型匹配，在黑麦培养基上产生卵孢子的量最大。越冬后卵孢子的存活率为19.57%~24.59%、萌发率为3.58%~8.49%。在全黑暗条件和光照黑暗交替条件下，卵孢子的萌发差异不明显。而土壤浸渍液比蒸馏水更适合培养卵孢子。
5.利用PCR-RFLP方法对甘肃甘肃省马铃薯晚疫病菌的mtDNA单倍型进行了分析，结果表明甘肃省马铃薯晚疫病菌的mtDNA单倍型为Ia和IIa，未发现Ib和IIb。Ia单倍型菌株分布在A1，A2和SF自育型交配型，分别占被测菌株的28.0%、40.0%和32.0%，90%IIa单倍型菌株为A1交配型。马铃薯晚疫病菌生理小种与mtDNA单倍型无明显的相关性。Ia单倍型菌株均对甲霜灵表现抗性。
6.对21个马铃薯品种（系）进行了田间抗性鉴定，结果表明，国瑞1号、临夏0308-2、0527-4、天水05-5-49-2和天水02-3-4对马铃薯晚疫病均表现高抗；天水05-7-5、天水06-3-4、陇薯6号、定西P2和27-7对马铃薯晚疫病表现均中抗；陇薯3号、中薯02175-200、0528-3、庄浪05-7-1和9901-10对马铃薯晚疫病表现中抗至感病；庄浪03-5-7、农大GDF、定西05A-1、LK99、生技所03-1和爱兰1号对马铃薯晚疫病均表现感病。
7.选用8个不同生理小种菌株，对甘肃省主栽品种及新品系进行室内抗病性鉴定，结果表明陇薯7号、陇薯8号、庄薯3号和青薯9号对所用马铃薯晚疫病菌表现抗性，陇薯3号、陇薯5号、陇薯9号、陇薯11号对部分菌株表现感病。
8．田间药剂筛选试验表明，亚磷酸-氢氧化钠盐病前500倍液喷雾对马铃薯晚疫病有较好的预防作用，防效可达68.00%～74.59%；60%氟吗啉·锰锌可湿性粉剂、50%氟啶胺悬浮剂和DPX-QGU42悬浮剂对马铃薯晚疫病的防治效果明显，防效分别为91.45%、88.32%、85.01%；生防菌剂寡雄腐霉可湿性粉剂防效达73.76%，而农用微生物菌剂枯草芽孢杆菌防效仅为17.50%～43.78%。
9．依据CARAH模型建立了病害预警预报技术，在马铃薯晚疫病的防控实践中加以运用；分析制定出适合甘肃省马铃薯晚疫病治理技术，为管理部门进行决策及生产部门实际操作提供了技术支撑。
Potato is dominant industry of agriculture in Gansu province. The potato lateblight disease caused by Phytophthora infestans de Bary occurs frequently, and leadsto serious damage in main potato area. The prevalence of the disease is determined byphenotypic structure of P. infestans, cultivar resistance and environmental factors. Inthis study, the potato late blight disease is main research object, including phenotypicstructure of P. infestans, survival and germination of overwintering oospores in soil,field screening of pesticides, cultivar resistant determination, and field controldirected by disease forecasting system. This study provides theoretical foundation andtechnical support to formulate integrated management of the potato late blight disease.
1. This study used international differential host R1-R11with single dominantresistance gene to determine race of75isolates of P. infestans from differentecoregions of potato production in Gansu province, and45races were determined,and the race l.2.3.4.5.6.7.8.9.l0.11occurred the highest frequently, and whichaccounted for22.67%ratio of determined isolates, was dominant race in Gansuprovince.
2.186isolates of P. infestans were assessed for sensitivity to Metalaxyl and28.5%of the isolates were sensitive to Metalaxyl,15.6%of the isolates wereintermediate resistance to Metalaxyl, and55.9%of the isolates were high resistance toMetalaxyl. The high resistance isolates to Metalaxyl were found in different collectingplaces, and showed that the isolates resistance of P.infestans to Metalaxyl were highfrequency in Gansu province.
3.147isolates mating types of P. infestans were determined by standard A1mating type and A2mating type. The result showed that there were4mating types ofP. infestans, such as A1mating type, A2mating type, A1A2mating type and SFmating type, which accounted for66.2%,20.4%,2.1%and16.3%, respectively.
4. Survival and germination of overwintering oospores of P. infestans found thatcompatible mating strains of P. infestans A1and A2produce oospores abundantly inpaired cultures on tomato-rye agar medium.The survival rate of overwintering wasranged from19.57%to24.59%.The germination rate of overwintering was rangedfrom3.58%to8.49%, and there were no significant differences in germination rate ofoospores between alternating light at day/night and whole darkness at day/night.Furthermore, the soil extract liquid was more suitable for oospore germination thandistilled water.
5. mtDNA haplotype of P. infestans in Gansu were analyzed, the result showedthat their mtDNA haplotype were Ia and IIa, not Ib and IIb. The isolates of Iahaplotype were distributed in A1mating type, A2mating type and SF mating type,and accounted for28.00%,40.00%, and32.00%of determined isolates, respectively.90.00%ratio of IIa haplotype was A1mating types. There was not significantrelevance between race and mtDNA haplotype of P. infestans. The isolates of Iahaplotype were resistant to Metalaxyl.
6. The field resistance of21cultivars (lines) was identified, and the resultshowed that the cultivar of Guorui-1, Lingxia0308-2,0527-4, Tianshui05-5-49-2and02-3-4was high resistance to P. infestans, the cultivar of Tianshui05-7-5,Tianshui6-3-4, Long6, P2and27-7was intermediate resistance to P. infestans, thecultivar of Long3, Zhongshu02175-200,0528-3, Zhuanglang05-7-1and9901-10wasintermediate resistance to susceptibility, the cultivar of Zhuanglang03-5-7,NongdaGDF, Dingxi05A-1, LK99, Shengjisuo03-1and Ailan1was susceptible to P.infestans.
7The result of cultivar resistant identification in Lab showed that the cultivar ofLong7, Long8, Zhuangshu3and Qingshu9was resistant to all race of P. infestans.Therefore, the cultivar of Long3, Long5, Long9, and Long11was susceptible to partisolate of P. infestans.
8The results of screening pesticides in field showed that500dilution water ofphosphite had perfect control effect to potato late blight, the control effect was68.00%-74.59%. The control effect of60%Flumorph mancozeb WP,50%FluazinamSC and DPX-QGU42SC was outstanding to potato late blight, the control effect was91.45%,88.32%,85.01%, respectively. Microbial agent Pythium Oligandrum WP was73.76%, however, for agricultural microbial agent Bacillus subtilis WP, the controleffect was only17.50%-43.78%.
9Disease forecasting technical system was established by CARAH method, andwhich conducted successfully disease control of potato late blight in field. Thatformulating integrated management of the potato late blight provided technicalsupport for decision of administration in Gansu province.
1.采用国际已知含单显性抗病基因系列品系R1-R11对采自甘肃马铃薯不同生态种植区的75个菌株进行了生理小种的鉴定，共鉴定出45个生理小种,其中生理小种l.2.3.4.5.6.7.8.9.l0.11出现频率最高，占被测菌株总数的22.67％，是甘肃省的优势生理小种。
2．对186个菌株进行甲霜灵敏感性测定，28.5％对甲霜灵表现敏感、15.6％表现中抗和55.9％表现高抗。各采集点均发现对甲霜灵表现高抗的菌株，这表明甘肃省马铃薯晚疫病菌对甲霜灵产生抗药性的频率比较高。
3.利用已知的A1、A2交配型标准菌株，对147个晚疫病菌株的交配型进行了测定，发现甘肃省马铃薯晚疫病菌交配型有A1、A2、A1A2、SF（自育型）4类，分别占被测菌株的66.2%、20.4%、2.1%、16.3%。
4.病原菌卵孢子存活和萌发试验发现在A1和A2交配型匹配，在黑麦培养基上产生卵孢子的量最大。越冬后卵孢子的存活率为19.57%~24.59%、萌发率为3.58%~8.49%。在全黑暗条件和光照黑暗交替条件下，卵孢子的萌发差异不明显。而土壤浸渍液比蒸馏水更适合培养卵孢子。
5.利用PCR-RFLP方法对甘肃甘肃省马铃薯晚疫病菌的mtDNA单倍型进行了分析，结果表明甘肃省马铃薯晚疫病菌的mtDNA单倍型为Ia和IIa，未发现Ib和IIb。Ia单倍型菌株分布在A1，A2和SF自育型交配型，分别占被测菌株的28.0%、40.0%和32.0%，90%IIa单倍型菌株为A1交配型。马铃薯晚疫病菌生理小种与mtDNA单倍型无明显的相关性。Ia单倍型菌株均对甲霜灵表现抗性。
6.对21个马铃薯品种（系）进行了田间抗性鉴定，结果表明，国瑞1号、临夏0308-2、0527-4、天水05-5-49-2和天水02-3-4对马铃薯晚疫病均表现高抗；天水05-7-5、天水06-3-4、陇薯6号、定西P2和27-7对马铃薯晚疫病表现均中抗；陇薯3号、中薯02175-200、0528-3、庄浪05-7-1和9901-10对马铃薯晚疫病表现中抗至感病；庄浪03-5-7、农大GDF、定西05A-1、LK99、生技所03-1和爱兰1号对马铃薯晚疫病均表现感病。
7.选用8个不同生理小种菌株，对甘肃省主栽品种及新品系进行室内抗病性鉴定，结果表明陇薯7号、陇薯8号、庄薯3号和青薯9号对所用马铃薯晚疫病菌表现抗性，陇薯3号、陇薯5号、陇薯9号、陇薯11号对部分菌株表现感病。
8．田间药剂筛选试验表明，亚磷酸-氢氧化钠盐病前500倍液喷雾对马铃薯晚疫病有较好的预防作用，防效可达68.00%～74.59%；60%氟吗啉·锰锌可湿性粉剂、50%氟啶胺悬浮剂和DPX-QGU42悬浮剂对马铃薯晚疫病的防治效果明显，防效分别为91.45%、88.32%、85.01%；生防菌剂寡雄腐霉可湿性粉剂防效达73.76%，而农用微生物菌剂枯草芽孢杆菌防效仅为17.50%～43.78%。
9．依据CARAH模型建立了病害预警预报技术，在马铃薯晚疫病的防控实践中加以运用；分析制定出适合甘肃省马铃薯晚疫病治理技术，为管理部门进行决策及生产部门实际操作提供了技术支撑。
Potato is dominant industry of agriculture in Gansu province. The potato lateblight disease caused by Phytophthora infestans de Bary occurs frequently, and leadsto serious damage in main potato area. The prevalence of the disease is determined byphenotypic structure of P. infestans, cultivar resistance and environmental factors. Inthis study, the potato late blight disease is main research object, including phenotypicstructure of P. infestans, survival and germination of overwintering oospores in soil,field screening of pesticides, cultivar resistant determination, and field controldirected by disease forecasting system. This study provides theoretical foundation andtechnical support to formulate integrated management of the potato late blight disease.
1. This study used international differential host R1-R11with single dominantresistance gene to determine race of75isolates of P. infestans from differentecoregions of potato production in Gansu province, and45races were determined,and the race l.2.3.4.5.6.7.8.9.l0.11occurred the highest frequently, and whichaccounted for22.67%ratio of determined isolates, was dominant race in Gansuprovince.
2.186isolates of P. infestans were assessed for sensitivity to Metalaxyl and28.5%of the isolates were sensitive to Metalaxyl,15.6%of the isolates wereintermediate resistance to Metalaxyl, and55.9%of the isolates were high resistance toMetalaxyl. The high resistance isolates to Metalaxyl were found in different collectingplaces, and showed that the isolates resistance of P.infestans to Metalaxyl were highfrequency in Gansu province.
3.147isolates mating types of P. infestans were determined by standard A1mating type and A2mating type. The result showed that there were4mating types ofP. infestans, such as A1mating type, A2mating type, A1A2mating type and SFmating type, which accounted for66.2%,20.4%,2.1%and16.3%, respectively.
4. Survival and germination of overwintering oospores of P. infestans found thatcompatible mating strains of P. infestans A1and A2produce oospores abundantly inpaired cultures on tomato-rye agar medium.The survival rate of overwintering wasranged from19.57%to24.59%.The germination rate of overwintering was rangedfrom3.58%to8.49%, and there were no significant differences in germination rate ofoospores between alternating light at day/night and whole darkness at day/night.Furthermore, the soil extract liquid was more suitable for oospore germination thandistilled water.
5. mtDNA haplotype of P. infestans in Gansu were analyzed, the result showedthat their mtDNA haplotype were Ia and IIa, not Ib and IIb. The isolates of Iahaplotype were distributed in A1mating type, A2mating type and SF mating type,and accounted for28.00%,40.00%, and32.00%of determined isolates, respectively.90.00%ratio of IIa haplotype was A1mating types. There was not significantrelevance between race and mtDNA haplotype of P. infestans. The isolates of Iahaplotype were resistant to Metalaxyl.
6. The field resistance of21cultivars (lines) was identified, and the resultshowed that the cultivar of Guorui-1, Lingxia0308-2,0527-4, Tianshui05-5-49-2and02-3-4was high resistance to P. infestans, the cultivar of Tianshui05-7-5,Tianshui6-3-4, Long6, P2and27-7was intermediate resistance to P. infestans, thecultivar of Long3, Zhongshu02175-200,0528-3, Zhuanglang05-7-1and9901-10wasintermediate resistance to susceptibility, the cultivar of Zhuanglang03-5-7,NongdaGDF, Dingxi05A-1, LK99, Shengjisuo03-1and Ailan1was susceptible to P.infestans.
7The result of cultivar resistant identification in Lab showed that the cultivar ofLong7, Long8, Zhuangshu3and Qingshu9was resistant to all race of P. infestans.Therefore, the cultivar of Long3, Long5, Long9, and Long11was susceptible to partisolate of P. infestans.
8The results of screening pesticides in field showed that500dilution water ofphosphite had perfect control effect to potato late blight, the control effect was68.00%-74.59%. The control effect of60%Flumorph mancozeb WP,50%FluazinamSC and DPX-QGU42SC was outstanding to potato late blight, the control effect was91.45%,88.32%,85.01%, respectively. Microbial agent Pythium Oligandrum WP was73.76%, however, for agricultural microbial agent Bacillus subtilis WP, the controleffect was only17.50%-43.78%.
9Disease forecasting technical system was established by CARAH method, andwhich conducted successfully disease control of potato late blight in field. Thatformulating integrated management of the potato late blight provided technicalsupport for decision of administration in Gansu province.
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