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Molecular Characterization, Morphological Characteristics, Virulence, and Geographic Distribution of Rhizoctonia spp. in Washington State

机译:根瘤菌属的分子特征,形态特征,毒力和地理分布在华盛顿州

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Rhizoctonia root rot and bare patch, caused by Rhizoctonia solani anastomosis group (AG)-8 and R. oryzae, are chronic and important yield-limiting diseases of wheat and barley in the Inland Pacific Northwest (PNW) of the United States. Major gaps remain in our understanding of the epidemiology of these diseases, in part because multiple Rhizoctonia AGs and species can be isolated from the same cereal roots from the field, contributing to the challenge of identifying the causal agents correctly. In this study, a collection totaling 498 isolates of Rhizoctonia was assembled from surveys conducted from 2000 to 2009, 2010, and 2011 over a wide range of cereal production fields throughout Washington State in the PNW. To determine the identity of the isolates, PCR with AG- or species specific primers and/or DNA sequence analysis of the internal transcribed spacers was performed. R. solani AG-2-1, AG-8, AG-10, AG-3, AG-4, and AG-11 comprised 157 (32%), 70 (14%), 21 (4%), 20 (4%), 1 (0.2%), and 1 (0.2%), respectively, of the total isolates. AG-I-like binucleate Rhizoctonia sp. comprised 44 (9%) of the total; and 53 (11%), 80 (16%), and 51 (10%) were identified as R. oryzae genotypes I, II, and III, respectively. Isolates of AG-2-1, the dominant Rhizoctonia, occurred in all six agronomic zones defined by annual precipitation and temperature within the region sampled. Isolates of AG-8 also were cosmopolitan in their distribution but the frequency of isolation varied among years, and they were most abundant in zones of low and moderate precipitation. R. oryzae was cosmopolitan, and collectively the three genotypes comprised 37% of the isolates. Only isolates of R. solani AG-8 and R. oryzae genotypes II and III (but not genotype I) caused symptoms typically associated with Rhizoctonia root rot and bare patch of wheat. Isolates of AG-2-1 caused only mild root rot and AG-I-like binucleate isolates and members of groups AG-3, AG-4, and AG-11 showed only slight or no discoloration of the roots. However, all isolates of AG-2-1 caused severe damping-off of canola, resulting in 100% mortality. Isolates of Rhizoctonia AG-8, AG-2-1, AG-10, AG-I-like binucleate Rhizoctonia, and R. oryzae genotypes I, II, and III could be distinguished by colony morphology on potato dextrose agar, by PCR with specific primers, or by the type and severity of disease on wheat and canola seedlings, and results of these approaches correlated completely. Based on cultured isolates, we also identified the geographic distribution of all of these Rhizoctonia isolates in cereal-based production systems throughout Washington State.
机译:Solsol solani吻合组(AG)-8和R. oryzae引起的Rhizoctonia根腐烂和裸露斑块是美国内陆西北太平洋(PNW)的小麦和大麦的慢性且重要的限产性疾病。在我们对这些疾病的流行病学的理解中,主要的差距仍然存在,部分原因是可以从田间的同一谷物根中分离出多种根瘤菌AG和菌种,这对正确确定病因进行了挑战。在这项研究中,从2000年至2009年,2010年和2011年对PNW华盛顿州整个谷物生产领域进行的调查中,收集了总计498株根瘤菌分离株。为了确定分离物的身份,进行了AG或物种特异性引物的PCR和/或内部转录间隔区的DNA序列分析。茄红霉菌AG-2-1,AG-8,AG-10,AG-3,AG-4和AG-11分别为157(32%),70(14%),21(4%),20(分别占总分离物的4%,1(0.2%)和1(0.2%)。类似于AG-I的双核根瘤菌。占总数的44(9%);分别将53和11(11%),80(16%)和51(10%)分别定为稻米红霉菌基因型I,II和III。 AG-2-1的分离株(主要的根瘤菌)出现在所有六个农艺区中,这六个农艺区由年降水量和采样区域内的温度确定。 AG-8分离株的分布也具有世界性,但分离的频率随年份变化,并且在低降水量和中等降水量地区最为丰富。米曲霉是世界性的,三种基因型共同构成了分离株的37%。仅R. solani AG-8和R. oryzae基因型II和III的分离株(而非基因型I)引起的症状通常与根瘤菌根腐病和小麦裸露斑有关。 AG-2-1的分离株仅引起轻微的根腐病和AG-1样的双核分离株,AG-3,AG-4和AG-11组的成员仅显示出轻微的根色或没有根色。但是,AG-2-1的所有分离物均导致双低油菜籽的严重减毒,导致100%的死亡率。可以通过马铃薯葡萄糖琼脂上的菌落形态学,通过PCR和PCR的方法来区分根瘤菌AG-8,AG-2-1,AG-10,AG-1样双核根霉和米曲霉基因型I,II和III的分离株。特定的引物,或小麦和油菜幼苗的病害类型和严重程度,这些方法的结果完全相关。基于培养的分离株,我们还确定了整个华盛顿州基于谷物的生产系统中所有这些根瘤菌分离株的地理分布。

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