Identification and use of a wild plant with antimicrobial activity against Ralstonia solanacearum, the cause of bacterial wilt of potato

Fresh aerial tissue and roots of 14 wild plants in Okinawa prefecture were investigated for their antimicrobial activity against Ralstonia solanacearum , which causes bacterial wilt of potato. A 70% aqueous ethanol extract of fresh aerial tissue of Geranium carolinianum L. showed strong antimicrobial activity against R. solanacearum . This extract also showed antimicrobial activity against the pathogens causing common scab of potato and soil rot of sweet potato. The antimicrobial substance could be extracted with hot water, and was effective against R. solanacearum in soil. In the field test, a treatment combining incorporation of dried aerial tissue into the soil and solarization was highly effective for control of bacterial wilt of potato. These findings suggest that G. carolinianum L. could be used as a biological agent for the control of bacterial wilt of potato.     

广东省烟草青枯茵的菌系和遗传多样性

从广东省韶关和梅州2个烟草产区采集了来自8个县市的共38个烟草青枯菌菌株，对病菌的生物型、致病型和遗传多样性进行了测定，结果表明广东烟草青枯菌全部为生物型Ⅲ。通过在红花大金元，K326，岩烟97和系3等4个烟草鉴别品种上的致病性反应，将广东烟草青枯菌分为强致病力、中等致病力和弱致病力3种致病型，其中以中等致病力菌株为主，占78．9％；强致病力菌株占13．2％；弱致病力菌株占7．9％。从116个RAPD通用引物中筛选出10条引物，用于对38个烟草青枯菌株DNA的RAPD分析，扩增条带表现出明显的多态性，条带主要聚集在0．3～4．0kb之间。聚类分析这38个菌株可分为2个组群，即组群A和组群B。组群A中又可以分为7个亚组（A1、A2、A3、A4、A5、A6、A7）；组群B也含有4个亚组（B1、B2、B3、B4）。研究结果还表明，广东烟草青枯菌菌株RAPD组群的划分与菌株的寄主来源有一定的相关性，但与地理区域、生物型和致病型没有明显的相关性；生物型不能反映烟草青枯菌菌系的差异。 04—0463—06     

土霉素对柑橘黄龙病的防治效果及对柑橘产量和品质的影响

为有效防控柑橘黄龙病,于2017年在佛罗里达大学柑橘研究与教育中心的奥本代尔市柑橘试验场进行田间试验筛选黄龙病的防治药剂及其浓度,测定注射土霉素后叶片中柑橘黄龙病菌亚洲种Cadidatus Liberibacter asiaticus、土霉素、淀粉含量、柑橘产量、出汁率、可溶性固形物含量和酸度,显微镜下观察注射土霉素后淀粉粒的分布。结果表明,浓度为1.6g/株土霉素处理180 d后柑橘叶片中黄龙病菌亚洲种含量减少幅度最大,为91.78%;注射浓度为1.6g/株土霉素后7d,叶片中土霉素含量达142.2 μg/kg,注射后11 d叶片中土霉素含量达到最大,为239.8 μg/kg,注射后45 d叶片中土霉素含量低至99.6 μg/kg以下;注射后7~90d,叶片中黄龙病菌亚洲种含量呈波浪形变化,注射90 d后叶片中黄龙病菌亚洲种含量一直处于增长趋势,叶片中黄龙病菌亚洲种含量总体随着土霉素含量的升高而降低;注射浓度为1.6g/株土霉素后,叶片中淀粉含量大幅度下降,60d时达到最低值,为3.3 μg/mm²,90d时达到峰值,为14.5 μg/mm²;单株产量为16.7 kg,与注射浓度为0.8 g/株土霉素处理差异不显著,但均显著高于其它2个处理;柑橘出汁率、可溶性固形物和酸度均与清水对照差异不显著。表明土霉素可有效抑制黄龙病菌亚洲种,减少柑橘叶片内淀粉含量,增加柑橘产量,但对果实品质未产生显著影响。     

Occurrence of prophage and historical perspectives associated with the dissemination of huanglongbing in mainland China

Huanglongbing (HLB), associated with a non-culturable bacterium ‘Candidatus Liberibacter asiaticus’ (CLas), is a highly destructive citrus disease with a long but poorly documented history in China. No effective treatment for HLB is available. The identification of new prophages in abundant CLas genomic sequence data provides new insights into both the diversity of CLas strains and HLB management. In this study, CLas populations from nine provinces were surveyed for the presence of prophage. Two major prophage typing groups (PTGs) were discovered to be associated with two different altitude regions: strains of CLas in PTG1 from high altitude regions (HAR) mainly contained prophage Type 1 only or Types 1 and 3, whereas strains of CLas in PTG2 from low altitude regions (LAR) mainly contained prophage Type 2. The discovery of these CLas population patterns provides evidence for independent origins of HLB in HAR and LAR. Guangdong province is the generally recognized domestic region of origin for HLB and is primarily responsible for the dissemination of HLB in LAR through transport of seedlings. Both Yunnan and Sichuan provinces are the probable regions of origin for HLB in HAR. PTG2 was further divided into two subgroups: PTG2-1, found in Guangdong, Fujian and Guangxi and PTG2-2, found in Jiangxi, Zhejiang and Hunan. These regions and prophage types are correlated with early and late introductions of HLB in LAR. These molecular analyses were supported by studying the history of the dissemination of HLB in historical documents.     

广东梅州柑橘黄龙病和病毒病的发生调查及其黄龙病菌原噬菌体多样性

【目的】调查柑橘黄龙病（Huanglongbing,HLB）和病毒病在广东省梅州市柑橘主产区的危害情况,分析该地区柑橘黄龙病菌（CLas）的原噬菌体多样性。【方法】以16S rDNA为模板设计引物,利用实时荧光定量PCR（qPCR)检测梅州市不同抽检地在2017—2019年3年间柑橘黄龙病的发病情况;同时分别采用已报道的柑橘衰退病毒（Citrus tristeza virus,CTV）、柑橘裂皮病毒（Citrus exocortis viroid,CEVd）、柑橘褪绿矮缩病毒（Citrus chlorotic dwarf-associated virus,CCDaV）、柑橘碎叶病毒（Citrus tatter leaf virus,CTLV）和柑橘黄脉病毒（Citrus yellow vein clearing virus,CYVCV）的特异性检测引物,提取样品的RNA,反转后以样品的cDNA为模板,通过普通PCR的方法分析梅州市抽检地区柑橘病毒病的发生情况。此外,以基于2种原噬菌体类型（SC1和SC2）对应的超变异基因区域设计的2对引物（Lap-TJ-F/Lap-TJ-R1和Lap-TJ-F/Lap-TJ-R2）,运用普通PCR方法分析该地区CLas菌株原噬菌体的遗传多样性。【结果】除2018年9月梅州大浦顺兴公司蜜柚基地的抽检样品外,梅州市其他被检地区均发现CLas。总体来说,梅州市的脐橙CLas检出率为55.3%,蜜柚为61.5%,沙田柚为61.7%。其中2017年采集自平原县大拓镇的蜜柚和沙田柚样品CLas检出率为100%,兴宁市白马镇的蜜柚为100%,沙田柚为80%;其他抽检地区的样品CLas检出率在16.7%—83.3%。虽然梅州市部分地区的检出率较高,但其病原菌的含量并不高,大部分处于柑橘黄龙病发病初、中期,属可防可控阶段;此外,梅州市柑橘病毒的总检出率不高,CTV、CTLV和CYVCV为梅州的主要病毒,CEVd和CCDaV没有检测到。CTV、CTLV和CYVCV在脐橙上的检出率依次为78.9%、7.9%和21.1%,蜜柚为15.4%、25.0%和9.6%,沙田柚为6.4%、2.1%和4.3%。基于Lap-TJ-F/Lap-TJ-R1和Lap-TJ-F/Lap-TJ-R2来研究CLas原噬菌体多样性的PCR扩增条带共有4种类型（SC1-1、SC1-2、SC2-1和SC2-2）,经过PCR电泳和测序得出,来自梅州市脐橙和沙田柚的CLas菌株以SC2-1型为主,蜜柚上以SC1-1型为主。【结论】梅州柑橘产区的柑橘黄龙病目前属于可防可控阶段,其病原菌菌株的原噬菌体在不同品种上具有其独特性;由于在抽检时发现有柑橘病毒病的存在,因此在种植过程中需加强种苗监管,同时应采取有效措施对柑橘黄龙病和柑橘木虱（Diaphorina citri）进行绿色防控。     

Effect of calcium concentration in nutrient solution before and after inoculation with Ralstonia solanacearum on resistance of tomato seedlings to bacterial wilt

We previously reported that calcium (Ca) nutrition in tomato (Lycopersicon esculentum Mill.) significantly affected the resistance to bacterial wilt caused by Ralstonia solanacearum Smith. To elucidate the mechanisms underlying the Ca-dependent resistance, the effect of the Ca concentration in the nutrient solution applied before and after inoculation with the pathogen on the resistance of tomato seedlings to bacterial wilt was studied. One week before inoculation, seedlings were transferred to nutrient solutions containing Ca at concentrations of 0.4, 4.4, or 20.4 mM. Soon after inoculation, the seedlings that were treated with each concentration of Ca before inoculation were transferred to solutions containing the same three concentrations of Ca. Although the disease development was not affected by the concentration of Ca in the solution before inoculation, a higher concentration of Ca after inoculation reduced the disease severity. This result suggests that the concentration of Ca in the host, especially in the cell walls, before infection may not be directly involved in the Ca-dependent resistance of tomato seedlings to bacterial wilt.     

青枯劳尔氏菌潜在新寄主鉴定与青枯病防治策略的思考

为了充分发挥农业措施管理,合理轮套作技术在防治农作物青枯病的作用,结合云南当地农业生态实际情况,采用分离自雪莲果青枯病植株上的青枯劳尔氏菌,在28℃温室条件下,遵循柯赫氏法则,接种、分离、再接种和再分离该病菌,完成了植物病害验证过程。结果表明：在接种后,杜氏鼠尾草、曼陀罗和鸭趾草等3种昆明地区田间常见杂草均出现青枯病典型症状,首先在接种部位出现黑色病斑,继而植株萎蔫,并有大量的乳白色菌脓溢出。结合在自然界下尚未发现这3种杂草感染青枯病和在根部有青枯劳尔氏菌富集的研究结果,这3种杂草被推断为青枯劳尔氏菌的潜在寄主。本研究结果对农作物青枯病防治策略有重要指导意义。     

芽孢杆菌在促进番茄生长和控制青枯病上的比较优势

在离体条件下对47个芽孢杆菌分离物进行了抑制番茄青枯病菌测定,其中12个表现良好的抑菌能力,通过细菌学和Biolog分析将其中5个代表性拮抗菌分别鉴定为枯草芽孢杆菌,解淀粉芽孢杆菌和地衣芽孢杆菌,其中枯草芽孢杆菌菌株的抑菌能力最强,解淀粉芽孢杆菌和地衣芽孢杆菌菌株其次.同时在温室条件下测定这5个芽孢杆菌拮抗菌促进番茄生长和诱导番茄对青枯病的抗性效果.用芽孢杆菌处理番茄种子能够有效地控制番茄青枯病,通过测定种子萌发,植株生长高度以及鲜重和干重表明芽孢杆菌的大多数分离物能够促进植物生长,同时这5个芽孢杆菌拮抗菌比对照显著地减少了青枯病的发病率,然而, 促生和病害抑制的效果因不同的菌株而异.其中枯草芽孢杆菌拮抗菌株不仅是良好的促生剂,也是有效的抗性诱导剂,它能减少青枯病80%,充分显示了芽孢杆菌在番茄青枯病治理上的潜在重要性.     

番茄青枯菌分泌物中热稳定成分的毒性研究

研究了青枯细菌毒性菌株平板培养滤液中热稳定性成分对番茄组织培养物的影响。结果表明;ＰＣＦ中含有一类非酶,非胞外多糖的热稳定毒性物质,它对番茄叶片脱分化过程有抑制作用,抗病材料较感病材料敏感,另外,它对番茄试管生根也起抑制作用;ＥＰＳ没有上述抑制作用。     

福建省花生青枯病菌致病型及生物型的测定

从采自福建 10个县 (市 )花生青枯病株标样中 ,分离出花生青枯病原菌菌株 72个 .对其中有代表性的15个菌株进行致病型测定 ,2 9个菌株进行生理生化测定 .供试菌株在鉴别寄主上的致病反应表明 ,福建省的花生青枯病菌属于致病型 和 ,其中以致病型 为主 .对乳糖、麦芽糖、纤维二糖、甘露醇、山梨醇和甜醇的利用能力以及对硝酸盐的还原作用表明 ,除晋江菌株的硝酸盐还原作用不产生气泡 ,定为生物型 - 4 ,同安菌株由于不利用三糖而利用三醇定为生物型 外 ,供测的其他菌株都属于生物型