阿月浑子炭疽病的鉴定及其防治
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摘要
本试验首次系统的对阿月浑子炭疽病的病原学、生物学特性和防治技术进行了研究。为阿月浑子炭疽病的生物防治和和化学防治打下了基础,主要研究结果如下:
1阿月浑子炭疽病菌的形态鉴定及ITS序列分析
本试验对阿月浑子炭疽病在河北省的危害、症状特点及病原菌的鉴定进行了研究。此病可危害叶片、果实和枝干,在叶、果上产生黑褐色病斑,在枝干上产生的病斑凹陷并流出透明状胶体。2005~2008年分别从河北涉县和河北唐县两栽培区采集发病材料,对病菌进行分离,获得22个菌落形态相同的菌株。这些菌株在PDA平板上25℃培养3~4 d后均长出灰白色、绒毛状菌落,5~7 d开始产生明显的桔红色粘质孢子团。此病害可引起草莓叶片发病,但对辣椒、葡萄、黄杨的叶片以及苹果的果实均不致病。分生孢子单胞,长椭圆形或圆筒形,两端钝圆,大小为7.9~13.2μm×3.3~4.8μm。附着胞深棕色,近圆形或不规则形,边缘整齐,壁厚,大小为3.6~4.8μm×4.2~6.4μm。以菌株CN504的DNA为模板,扩增得到了全长为583 bp的DNA片段,并获得了该菌的rDNA-ITS序列。将该序列与GenBank中已有的DNA序列进行同源性比较,发现与其同源性最高的100个ITS序列菌株均为胶孢炭疽菌Colletotrichum gloeosporioides Penz或胶孢炭疽菌的有性态围小丛壳菌Glomerella cingulata Stenem,CN504与它们的同源性为98%~99%。根据病原菌的形态和生物学特征,以及ITS序列同源性比较的结果,将阿月浑子炭疽病的病原菌鉴定为胶孢炭疽菌C. gloeosporioides。
2阿月浑子炭疽病的防治
本试验首次系统地对阿月浑子炭疽病防治技术进行了研究。研究了四种生防菌和八种药剂对阿月浑子炭疽病菌的抑制活性,抑制机制以及对阿月浑子炭疽病的防治效果。平板抑菌圈法试验结果表明,生防菌CN068、WCS358::phl对病原菌有较强的拮抗活性;药剂炭阻绝、炭疽福美和代森锰锌对病原菌有的抑制能力较强。室内防治结果表明CN068在品种T-491叶片上防治效果较好,防治率达87.5%,而WCS358::phl防治效果较差;药剂炭疽福美、代森锰锌防效最好。田间防治试验验证两种药剂分别单独使用和交替使用三处理对阿月浑子炭疽病的防治都有极显著效果(炭疽福美的平均防治率为44.30%,代森锰锌为38.93%,两药交替使用为42.87%)。其中炭疽福美的防治效果最为稳定,建议在以后的防治工作中推广使用。
3阿月浑子抗病品种的筛选
通过室内试验调查阿月浑子叶片接种的发病时间和发病面积可知品种T-491,新疆选优,N品种,M-38,T-662的抗病性较强,田间试验显示品种T-491,T-471和T-11的抗病性最强。综上认为T-491是抗病性最优的品种。
In this research, the pathogen, biological characteristics and disease control of anthracnose in Pistacia vera L. were investigated. The main results are as follows:
1 Identification and ITS sequence analysis of the pathogen of Pistacia vera L. anthracnose
The occurrence, symptom characteristics and pathogen identification of anthracnose on Pistacia vera L. in Hebei were studied. This disease caused damage on leaves, fruits and branch. There are brown spot on the leaves and fruits, and canker on branches with transparent exudate. Twenty two strains were collected from two districts from 2005 to 2008 and all isolates showed similar characteristics in colony. They formed gray villous colonies in PDA culture at 25℃after 3~4 d, and orange spore mass appeared after culturing for 5~7 d. This fungus could infect strawberry leaves, but not pepper, grape, Japan euonymus leaves and the fruits of apple. Conidium is long oval or cylindrical with a single cell, 7.9~13.2μm×3.3~4.8μm, blunt at both ends. Appressorium, 3.6~4.8μm×4.2~6.4μm, is dark brown, with nearly round or irregular shape and thick wall. A rDNA-ITS fragment of 583 bp was amplified using the DNA of strain CN504 as a template and sequenced. The obtained sequences was compared with those in GenBank, and we found that 100 ITS sequences with the highest homology belong to Colletotrichum gloeosporioides Penz or the sexual stage of Glomerella cingulata Stenem, and their homology is 98% ~ 99%. According to its characteristics of morphology, pathogenicity and sequence analysis of the internal transcribed spacer region of the ribosomal DNA (ITS), the pistachio pathogen was identified as C. gloeosporioides.
2 The Control of Anthracnose in Pistacia vera
The control measures of anthracnose in Pistacia vera L. were investigated for the first time in the research. Inhibition activity, mechanism and control effect of four antagonistic microorganism and eight fungicides to the pathogen of anthracnose in Pistacia vera were studied in detail. The results of control effect in vitro showed that CN068 had the strongest antagonistic activities in the leaves of variety T-491, WCS358::phl had the worse antagonistic activities. Ziram-thira and mancozeb had good effect. Results of control effect in the field shows that two fungicides and their mixture all had distinct control effect. Ziram-thira should be extend because of its best Stability in the control.
3 The selection of resistance varieties of Pistacia vera
In vitro inoculations were carried out in the laboratory, and the infection time and area were investigated .The results showed: T-471, T-491, Sinkiang-Best, Varieties-N, M-38, T-662 have the stronger resistance. The performance of field showed T-471, T-11 are strongest resistant varieties. Based on the above-mentioned test, variety T-471 is the most resistant one.
1阿月浑子炭疽病菌的形态鉴定及ITS序列分析
本试验对阿月浑子炭疽病在河北省的危害、症状特点及病原菌的鉴定进行了研究。此病可危害叶片、果实和枝干,在叶、果上产生黑褐色病斑,在枝干上产生的病斑凹陷并流出透明状胶体。2005~2008年分别从河北涉县和河北唐县两栽培区采集发病材料,对病菌进行分离,获得22个菌落形态相同的菌株。这些菌株在PDA平板上25℃培养3~4 d后均长出灰白色、绒毛状菌落,5~7 d开始产生明显的桔红色粘质孢子团。此病害可引起草莓叶片发病,但对辣椒、葡萄、黄杨的叶片以及苹果的果实均不致病。分生孢子单胞,长椭圆形或圆筒形,两端钝圆,大小为7.9~13.2μm×3.3~4.8μm。附着胞深棕色,近圆形或不规则形,边缘整齐,壁厚,大小为3.6~4.8μm×4.2~6.4μm。以菌株CN504的DNA为模板,扩增得到了全长为583 bp的DNA片段,并获得了该菌的rDNA-ITS序列。将该序列与GenBank中已有的DNA序列进行同源性比较,发现与其同源性最高的100个ITS序列菌株均为胶孢炭疽菌Colletotrichum gloeosporioides Penz或胶孢炭疽菌的有性态围小丛壳菌Glomerella cingulata Stenem,CN504与它们的同源性为98%~99%。根据病原菌的形态和生物学特征,以及ITS序列同源性比较的结果,将阿月浑子炭疽病的病原菌鉴定为胶孢炭疽菌C. gloeosporioides。
2阿月浑子炭疽病的防治
本试验首次系统地对阿月浑子炭疽病防治技术进行了研究。研究了四种生防菌和八种药剂对阿月浑子炭疽病菌的抑制活性,抑制机制以及对阿月浑子炭疽病的防治效果。平板抑菌圈法试验结果表明,生防菌CN068、WCS358::phl对病原菌有较强的拮抗活性;药剂炭阻绝、炭疽福美和代森锰锌对病原菌有的抑制能力较强。室内防治结果表明CN068在品种T-491叶片上防治效果较好,防治率达87.5%,而WCS358::phl防治效果较差;药剂炭疽福美、代森锰锌防效最好。田间防治试验验证两种药剂分别单独使用和交替使用三处理对阿月浑子炭疽病的防治都有极显著效果(炭疽福美的平均防治率为44.30%,代森锰锌为38.93%,两药交替使用为42.87%)。其中炭疽福美的防治效果最为稳定,建议在以后的防治工作中推广使用。
3阿月浑子抗病品种的筛选
通过室内试验调查阿月浑子叶片接种的发病时间和发病面积可知品种T-491,新疆选优,N品种,M-38,T-662的抗病性较强,田间试验显示品种T-491,T-471和T-11的抗病性最强。综上认为T-491是抗病性最优的品种。
In this research, the pathogen, biological characteristics and disease control of anthracnose in Pistacia vera L. were investigated. The main results are as follows:
1 Identification and ITS sequence analysis of the pathogen of Pistacia vera L. anthracnose
The occurrence, symptom characteristics and pathogen identification of anthracnose on Pistacia vera L. in Hebei were studied. This disease caused damage on leaves, fruits and branch. There are brown spot on the leaves and fruits, and canker on branches with transparent exudate. Twenty two strains were collected from two districts from 2005 to 2008 and all isolates showed similar characteristics in colony. They formed gray villous colonies in PDA culture at 25℃after 3~4 d, and orange spore mass appeared after culturing for 5~7 d. This fungus could infect strawberry leaves, but not pepper, grape, Japan euonymus leaves and the fruits of apple. Conidium is long oval or cylindrical with a single cell, 7.9~13.2μm×3.3~4.8μm, blunt at both ends. Appressorium, 3.6~4.8μm×4.2~6.4μm, is dark brown, with nearly round or irregular shape and thick wall. A rDNA-ITS fragment of 583 bp was amplified using the DNA of strain CN504 as a template and sequenced. The obtained sequences was compared with those in GenBank, and we found that 100 ITS sequences with the highest homology belong to Colletotrichum gloeosporioides Penz or the sexual stage of Glomerella cingulata Stenem, and their homology is 98% ~ 99%. According to its characteristics of morphology, pathogenicity and sequence analysis of the internal transcribed spacer region of the ribosomal DNA (ITS), the pistachio pathogen was identified as C. gloeosporioides.
2 The Control of Anthracnose in Pistacia vera
The control measures of anthracnose in Pistacia vera L. were investigated for the first time in the research. Inhibition activity, mechanism and control effect of four antagonistic microorganism and eight fungicides to the pathogen of anthracnose in Pistacia vera were studied in detail. The results of control effect in vitro showed that CN068 had the strongest antagonistic activities in the leaves of variety T-491, WCS358::phl had the worse antagonistic activities. Ziram-thira and mancozeb had good effect. Results of control effect in the field shows that two fungicides and their mixture all had distinct control effect. Ziram-thira should be extend because of its best Stability in the control.
3 The selection of resistance varieties of Pistacia vera
In vitro inoculations were carried out in the laboratory, and the infection time and area were investigated .The results showed: T-471, T-491, Sinkiang-Best, Varieties-N, M-38, T-662 have the stronger resistance. The performance of field showed T-471, T-11 are strongest resistant varieties. Based on the above-mentioned test, variety T-471 is the most resistant one.
引文
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