黑龙江省大豆品种对大豆疫霉根腐病抗性评价及抗性基因推导

采用下胚轴伤口接种法,利用课题组在黑龙江省分离鉴定的8个大豆疫霉根腐病生理小种（race1、race3、race4、race5、race9、race13、race44、race54）对黑龙江省曾经大面积推广种植的44个大豆品种进行接种鉴定。结果表明,对8个生理小种分别有45.5%、29.5%、40.9%、43.2%、36.8%、45.5%、31.8%、72.7%表现抗性或中间类型,共产生37种反应型;通过基因推导,合丰-44,绥农-12可能含有Rps1c基因或Rps1a＋Rps1c、Rps1c＋Rps6、Rps1c＋Rps7的基因组合;黑河-32可能含有Rps1a＋Rps1d的基因组合,其它品种可能含有新的抗病基因或基因组合,表明黑龙江省存在丰富的抗大豆疫霉根腐病的种质资源。     

Characteristic of the Pepper CaRGA2 Gene in Defense Responses against Phytophthora capsici Leonian

The most significant threat to pepper production worldwide is the Phytophthora blight, which is caused by the oomycete pathogen, Phytophthora capsici Leonian. In an effort to help control this disease, we isolated and characterized a P. capsici resistance gene, CaRGA2, from a high resistant pepper (C. annuum CM334) and analyzed its function by the method of real-time PCR and virus-induced gene silencing (VIGS). The CaRGA2 has a full-length cDNA of 3,018 bp with 2,874 bp open reading frame (ORF) and encodes a 957-aa protein. The protein has a predicted molecular weight of 108.6 kDa, and the isoelectric point is 8.106. Quantitative real-time PCR indicated that CaRGA2 expression was rapidly induced by P. capsici. The gene expression pattern was different between the resistant and susceptible cultivars. CaRGA2 was quickly expressed in the resistant cultivar, CM334, and reached to a peak at 24 h after inoculation with P. capsici, five-fold higher than that of susceptible cultivar. Our results suggest that CaRGA2 has a distinct pattern of expression and plays a critical role in P. capsici stress tolerance. When the CaRGA2 gene was silenced via VIGS, the resistance level was clearly suppressed, an observation that was supported by semi-quantitative RT-PCR and detached leave inoculation. VIGS analysis revealed their importance in the surveillance to P. capsici in pepper. Our results support the idea that the CaRGA2 gene may show their response in resistance against P. capsici. These analyses will aid in an effort towards breeding for broad and durable resistance in economically important pepper cultivars.     

烟草低头黑病菌毒素钝化物的初步研究

研究了10种化合物4个浓度对烟草低头黑病菌(Colletotrichum capsici f.nicotianae)毒素的钝化作用。结果表明，高锰酸钾、氢氧化钠和碳酸氢钠对毒素具有很强的钝化作用。高锰酸钾在离体叶片上的相对防效为100％；各浓度的氢氧化钠和碳酸氢钠均对烟草低头黑病菌菌丝生长和分生孢子萌发具有一定的抑制作用。高锰酸钾并不抑制菌丝生长。     

烟草黑胫病菌对甲霜灵的抗药性风险

为了评估烟草黑胫病菌对甲霜灵的抗药性风险,以对甲霜灵敏感和高抗的烟草黑胫病菌240和191为出发菌株,进行了抗性突变频率测定、抗性诱变、抗性水平测定、遗传稳定性、生长速率、致病性测定和适合度等研究。结果表明,烟草黑胫病菌对甲霜灵敏感的240菌株在含10mg/L甲霜灵的CA平板上的抗性突变频率为7.0×10-5。分别以药剂诱变和紫外诱变获得了抗甲霜灵菌株240-YY和240-ZY。对甲霜灵敏感性不同的烟草黑胫病菌适合度测定结果表明,抗性菌株在潜伏期、侵染率、病情指数和病斑面积等方面均具有较高的适合度水平。研究结果显示,烟草黑胫病菌对甲霜灵具有较大的抗药性风险。     

Genetic Diversity and Phylogeny of Antagonistic Bacteria against Phytophthora nicotianae Isolated from Tobacco Rhizosphere

The genetic diversity of antagonistic bacteria from the tobacco rhizosphere was examined by BOXAIR-PCR, 16S-RFLP, 16S rRNA sequence homology and phylogenetic analysis methods. These studies revealed that 4.01% of the 6652 tested had some inhibitory activity against Phytophthora nicotianae. BOXAIR-PCR analysis revealed 35 distinct amplimers aligning at a 91% similarity level, reflecting a high degree of genotypic diversity among the antagonistic bacteria. A total of 25 16S-RFLP patterns were identified representing over 33 species from 17 different genera. Our results also found a significant amount of bacterial diversity among the antagonistic bacteria compared to other published reports. For the first time; Delftia tsuruhatensis, Stenotrophomonas maltophilia, Advenella incenata, Bacillus altitudinis, Kocuria palustris, Bacillus licheniformis, Agrobacterium tumefaciens and Myroides odoratimimus are reported to display antagonistic activity towards Phytophthora nicotianae. Furthermore, the majority (75%) of the isolates assayed for antagonistic activity were Gram-positives compared to only 25% that were Gram-negative bacteria.     

大豆资源抗疫霉根腐病基因分析

采取下胚轴创伤接种法鉴定156份大豆资源对13个不同毒力基因型大豆疫霉菌株的抗性。结果表明,125份资源分别抗1～13个菌株,占鉴定资源总数的80.13%。125份抗性大豆资源与13个大豆疫霉菌株共产生90种反应型。通过与13个鉴别寄主的反应型比较发现,有9份大豆资源产生的5种反应型与含有已知抗病基因的大豆资源的反应型相同;12份大豆资源产生的5种反应型与已知2个抗病基因组合的反应型一致,另外,还有至少抗1个菌株的104份大豆资源产生的80种反应型,既不同于已知单个抗病基因的反应型,也不同于2个已知抗病基因组合的反应型,推测可能含有新的抗病基因或基因组合。