Superoxide anion and hydrogen peroxide in the yeast antagonist–fruit interaction: A new role for reactive oxygen species in postharvest biocontrol?

The importance of reactive oxygen species (ROS) in plant defense responses against certain pathogens is well documented. There is some evidence that microbial biocontrol agents also induce a transient production of ROS in a host plant which triggers local and systemic defense responses to pathogens. The ability of biocontrol agents used to control postharvest diseases to induce defense-related oxidative responses in fruits, however, has not been explored. Here we show that the yeast antagonists, Metschnikowia fructicola (strain 277) and Candida oleophila (strain 182) generate greater levels of super oxide anion (O₂⁻) on intact fruit surfaces (poor in nutrients) then those applied on a nutrient-poor agar medium. Even though yeast antagonists show a high level of O₂⁻ on nutrient-rich media, when applied on fruits around wounds (areas abundant in nutrients) accumulation of O₂⁻, as detected by nitro blue tetrazolium staining, occurred much more rapidly on the latter. Using laser scanning confocal microscopy we observed that the application of M. fructicola and C. oleophila into citrus and apple fruit wounds correlated with an increase in H₂O₂ accumulation in host tissue. In citrus fruit, the level of H₂O₂ around inoculated wounds increased by 4-fold compared to controls (wounds inoculated with water) as early as 18 h after inoculation. Yeast continued to stimulate H₂O₂ production in citrus fruit up to 66 h after inoculation and H₂O₂ levels were still 3-fold above the control. Living yeast cells were detected in fruit wounds at this time point indicating the ability of M. fructicola to tolerate host ROS, which has been reported to be an intrinsic characteristic of efficient yeast antagonists. Similar increase in H₂O₂ accumulation around yeast-inoculated wounds was observed in apple fruit exocarp. The present data, together with our earlier discovery of the importance of H₂O₂ production in the defense response of citrus flavedo to postharvest pathogens, indicate that the yeast-induced oxidative response in fruit exocarp may be associated with the ability of specific yeast species to serve as biocontrol agents for the management of postharvest diseases.     

利用基因YKT6鉴定疫霉属的不同种

 利用分子标记或对特异位点的碱基序列进行分析是植物病原物分子检测的基础,可以在属和种的水平上对物种进行区分和鉴定。对疫霉属的不同种已有一系列的分子检测方法。SNARE蛋白相关基因YKT6拥有保守的侧翼编码区,适于设计疫霉属特异性的PCR引物,同时其内含子所具有的多态性可开发出几乎所有疫霉种的分子标记。利用疫霉属特异性引物对P-YKT6-F/P-YKT6-R可在31个疫霉种中特异地扩增出一条约600 bp的条带,而在腐霉或其他真菌中不能扩增出该条带。利用大豆疫霉的引物对Ps-YKT6-F/ Ps-YKT6-R和辣椒疫霉的引物对Pc-YKT6-F/Pc-YKT6-R,能分别从大豆疫霉菌株和辣椒疫霉菌株中扩增出一条399 bp和282 bp的条带,常规PCR和巢式PCR的灵敏度分别达到100 pg和10 fg。利用这些引物也可从土壤和病组织中检测到目标病原菌。此外,利用上述特异性引物开发出了大豆疫霉和辣椒疫霉的实时定量PCR检测方法。基于YKT6基因的分子标记和检测方法可用于疫霉种的调查检测和法定定量检测。     

戊唑醇在葡萄和土壤中的残留和消解动态

建立了葡萄中戊唑醇残留的气相色谱测定方法,并研究了其在葡萄和土壤中的消解动态。土壤用乙腈提取,无需净化,葡萄样品用甲醇提取,二氯甲烷液液分配净化后用气相色谱-氮磷检测器(GC-NPD)测定。结果表明:在 0.01、0.1、1 mg/kg 3个添加水平下,戊唑醇的平均回收率为85.0%~98.8%,相对标准偏差(RSD)为2.9%~10.4%;最小检出量为1×10-11g,最低检测浓度为0.01 mg/kg。采用250 g/L戊唑醇水乳剂按有效成分187.5 mg/L剂量(推荐剂量的1.5倍)施药,戊唑醇在葡萄中的半衰期为9.8~12.2 d,在土壤中的半衰期为8.2~17.3 d,药后28、35 d葡萄中的最终残留量≤0.81 mg/kg,低于国际食品法典委员会(CAC)和中国规定的最大残留限量2.0 mg/kg。建议在葡萄上使用250 g/L戊唑醇水乳剂时,施药剂量最高为有效成分187.5 mg/L,施药2~3次,采收间隔期为28 d。     

Maintenance of broccoli quality and functional properties during cold storage as affected by modified atmosphere packaging

Broccoli (Brassica oleracea L. var. Italica) heads were packaged using 3 types of polypropylene films: macro-perforated (Ma-P), micro-perforated (Mi-P) and non-perforated (No-P), and then stored at 1 °C for 28 days to study the effects of modified atmosphere packaging (MAP) on the maintenance of quality and functional properties by comparison with non-wrapped heads. Results revealed that deterioration occurred quickly in control broccoli, manifested mainly by weight loss, yellowing, chlorophyll degradation and stem hardening. Also, a rapid decrease in total antioxidant activity (TAA), ascorbic acid and total phenolic compound concentration was observed. Conversely, in those heads packaged under MAP, especially for Mi-P and No-P, all changes related with loss of quality were significantly reduced and delayed with time. Additionally, TAA, ascorbic acid and total phenolic compounds remained almost unchanged during the whole period. Thus, broccoli packaged with Mi-P and No-P films had prolonged storability up to 28 days with high quality attributes and health-promoting compounds, this period being only 5 days in unwrapped control broccoli.     

基于COⅠ和Cytb基因的浙江不同抗性水平二化螟种群的遗传结构分析

本研究利用线粒体细胞色素氧化酶亚基Ⅰ(COⅠ)和细胞色素b(Cytb)基因的遗传学方法分析了浙江省8个不同抗性水平二化螟地理种群的遗传多样性及种群遗传结构。种群遗传多样性分析表明:PCR 扩增测序分别获得长度为627 bp的COⅠ基因片段和长度为455 bp的Cytb基因片段。355条同源COⅠ序列监测出了68个多态性位点,其中单突变位点22个,简约突变位点46个,共定义了85个单倍型,每个群体的平均单倍型为18.25个,其中瑞安(RA)种群中单倍型最多,为27个单倍型。326条Cytb同源序列监测出了45个多态性位点,其中单突变位点19个,简约突变位点26个,共定义了64个单倍型,每个群体的平均单倍型为14.375个,其中乐清(YQ)种群中单倍型最多,为25个单倍型。此外,各群体中最高的单倍型多样度h分别为0.896 3和0.934 4,反映出二化螟群体较低的遗传多样性水平。种群遗传结构分析表明:二化螟不同地理种群间遗传变异大多数来自于群体内个体间,占80.30%(COⅠ基因)和78.16%(Cytb基因)。较少部分的遗传差异来自于组间,仅占19.43%(COⅠ基因)和21.22%(Cytb基因)。单倍型网络关系图未表现出显著的地理谱系结构,Mantel相关性检测显示,遗传距离与地理距离之间无显著相关性。单倍型邻接树也没有明显分支,未呈现出地域性差异。该研究结果为浙江省不同抗性水平二化螟种群间交流和二化螟的防控提供了基础资料。     

黄瓜绿斑驳花叶病毒安徽分离物全基因组序列测定及分析

通过胶体金免疫层析试纸条和RT-PCR等手段对采自安徽和县的西瓜病株进行检测,确定其病原为黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus,CGMMV)。为明确CGMMV安徽分离物CGMMV-Anhui的分类地位,进一步克隆了该病毒的全基因组序列,分析了其基因组结构特征。结果表明,CGMMV-Anhui基因组全长为6 423bp(GenBank登录号KT236095),与已报道的CGMMV的编码区的基因结构一致,仅5′和3′端非编码区核苷酸数目略有差异。将CGMMV-Anhui与已报道的分离物的全基因组序列和外壳蛋白基因序列进行系统发育分析,显示CGMMV不同分离物可分为亚洲和欧洲两个组,安徽分离物CGMMV-Anhui与亚洲分离物亲缘关系较近,可能具有共同的侵染源。     

Ypt1 Gene-Based Detection of Phytophthora Sojae in a Loop-Mediated Isothermal Amplification Assay

Journal of Plant Diseases and Protection - This study reports the development of a loop-mediated isothermal amplification procedure (LAMP) for the detection of Phytophthora sojae, the causal agent...     

解淀粉芽胞杆菌WH1G与咪鲜胺协同防治草莓炭疽病

通过室内测定8种化学药剂和拮抗细菌解淀粉芽胞杆菌WH1G对草莓炭疽病菌的毒力,将毒力最强的化学药剂与WH1G进行复配,探讨复配剂对草莓炭疽病菌的室内毒力及田间防治效果。室内生物测定结果表明,8种常见化学药剂中咪鲜胺对草莓炭疽病菌的毒力最强,其EC50为0.045 3mg/L,在较低浓度下能有效抑制草莓炭疽病菌的生长,与生防菌有较好的生物相容性;解淀粉芽胞杆菌WH1G对草莓炭疽病菌菌丝生长具有较强的抑制作用,含菌量为1.0×1010cfu/mL时抑制率达89%;将咪鲜胺(0.045 3mg/L)与解淀粉芽胞杆菌WH1G(2.3×106cfu/mL)复配,V(咪鲜胺)∶V(WH1G)配比为5∶5时对病菌抑制的增效作用和防治效果最好,毒性比率为1.432,防效为69.94%。复配剂对草莓炭疽病的田间防效达67.91%,显著高于单剂防效,且咪鲜胺使用量只有单剂使用量的1/2,表明二者复配不仅可以提高防效,还能有效减少化学药剂的使用量。     

我国豇豆蓟马研究进展及综合防控措施

豇豆Vigna unguiculata 喜温喜光,是一种适合在我国南方种植的豆科蔬菜。长期以来,豇豆病虫害多发重发,其中,蓟马是豇豆上最主要和最顽固的害虫之一。传统的化学防治方法对蓟马的防治效果常常不理想,并易造成农药残留问题。本文概述了我国豇豆产业的种植情况和以蓟马为主的病虫害问题及豇豆蓟马研究现状,并从农艺措施、理化诱控、生物防治、化学防治和抗性治理角度总结了豇豆蓟马的综合防治技术。最后,从构建豇豆蓟马的有害生物综合治理(integrated pest management, IPM)技术体系、加强基础研究和防控技术产品开发、建立科学用药和农药监管制度3个方面提出建议,以期为我国豇豆产业的绿色可持续发展提供参考。     

安徽北部小麦田草地贪夜蛾发生情况及田间防治试验

2019年11月9日至15日对安徽北部小麦田草地贪夜蛾发生情况进行了调查,发现涡阳县、蒙城县、利辛县、谯城区、临泉县等地部分田块为害偏重,每平方米最高虫量达51头,百株被害率最高为66.85%。利用C OⅠ和Tpi基因进行遗传分析,发现采自涡阳小麦田的试虫均为玉米型。田间防治试验结果表明,10%甲维盐·氯虫苯甲酰胺悬浮剂和12%甲维盐·虫螨腈悬浮剂药后3 d防效分别为90.64%和92.17%;药后14 d防效分别为91.32%和92.26%,速效性和持效性皆较佳,可作为防治小麦田草地贪夜蛾的推荐药剂。