植物源与微生物源生物制剂复配防治根结线虫病

doi:10.13560/j.cnki.biotech.bull.1985.2021-0408

摘要/Abstract

摘要：

番茄根结线虫病严重影响农业生产,利用植物源和微生物源两种不同的生物制剂复配防治番茄根结线虫病,试图探索出适应绿色农业生产需求的一种新的可能性。通过稀释涂布平板法从土壤分离出3株12 h内对根结线虫二龄幼虫（second-stage juveniles,J2）抑杀率分别达89.23%、87.95%、88.97%的生防菌株SJ5、SJ17、SJ19,经16S rDNA扩增测序鉴定分别为耐冷假单胞菌（Pseudomonas psychrotolerans）、黏金黄杆菌（Chryseobacterium gleum）和枯草芽孢杆菌（Bacillus subtilis）。通过平板抑菌圈法证明3种生防菌之间、生防菌与苦参印楝素之间无拮抗现象,适宜复配。盆栽实验初步验证,发酵48 h的3种生防菌发酵液（有效活菌数≥3.0 亿个/mL）与苦参印楝素稀释液（1 000 倍）复配制剂处理组对根结线虫病的防效达67.75%,与化学农药（氟吡菌酰胺、噻唑膦）处理组相比防效无显著性差异,且番茄株高、茎围、苗和根部的生物量均显著性增加,盆栽试验结论初步发现复配方案具有防治番茄根结线虫病害应用潜力。

关键词: 番茄, 根结线虫, 生物防治, 复配

Abstract:

Tomato root-knot nematode disease seriously affects agricultural production. The compound of two biological agents from plant origin and microbial source are used to control tomato root-knot nematode disease,trying to explore a new feasibility to meet the need of green agricultural production. Three strains（SJ5,SJ17 and SJ19）of killing second-stage juveniles（J2）of root knot nematodes were isolated from the soil by the dilution coating plate method with an inhibitory rate of 89.23%,87.95%,and 88.97%,respectively,and they were identified as Pseudomonas psychrotolerans,Chryseobacterium gleum and Bacillus subtilis by 16S rDNA amplification and sequencing. The plate inhibition zone method proved that there was no antagonism among the three kinds of biocontrol bacteria,and between the biocontrol bacteria and azadirachtin,thus they were suitable for compounding. Preliminary verification by pot experiment showed that the compound preparation treatment group from the filtrates by three kinds of biocontrol bacteria fermented for 48 h（the effective number of viable bacteria in the fermentation broth was more than 300 million/mL）and the azadirachtin diluent（1 000 times）was effective against root-knot nematodes The control effect of the disease reached 67.75%,and there was no significant difference in the control effect compared with the chemical pesticides（fluopyram and thiazophos）treatment group,and the tomato’s plant height,stem circumference,seedling and root biomass were all significantly increased. The preliminary conclusion from the potted experiment reveals that the compound scheme has the application potential to control tomato root-knot nematode disease.

Key words: Solanum lycopersicum, root-knot nematode, biological control, compound

舒洁, 张仁军, 梁应冲, 陈雅琼, 张娟, 郭建, 陈穗云. 植物源与微生物源生物制剂复配防治根结线虫病[J]. 生物技术通报, 2021, 37(7): 164-174.

SHU Jie, ZHANG Ren-jun, LIANG Ying-chong, CHEN Ya-qiong, ZHANG Juan, GUO Jian, CHEN Sui-yun. Control of Root-knot Nematode Disease by Compounding Biological Agents from Plant and Microorganisms[J]. Biotechnology Bulletin, 2021, 37(7): 164-174.

图/表 10

图1 SJ5、SJ17、SJ19菌体形态

Fig.1 Morphology of strain SJ5,SJ17,and SJ19

图2 菌株SJ5系统发育树

Fig.2 Phylogenetic tree of SJ5

图3 菌株SJ17系统发育树

Fig.3 Phylogenetic tree of SJ17

图4 菌株SJ19系统发育树

Fig.4 Phylogenetic tree of SJ19

图5 显微镜下观察线虫毒力测定 ①LB对照;②SJ5发酵液;③SJ17发酵液;④SJ19发酵液;⑤苦参印楝素（1 000倍）。线虫虫体弯曲表示处于活体状态,弧形或直线形状代表已死亡或者僵直濒临死亡状态

Fig. 5 Virulence determination of nematodes under microscope ① LB control;② SJ5 fermentation filtrate;③ SJ17 fermentation filtrate;④ SJ19 fermentation filtrate;⑤ Sophora azadirachtin dilution（1 000 times）. The bent body of the nematode indicates that it is in a living state,and the arc or straight shape indicates that it is dead or stiff and dying

图6 不同处理组毒力测试差异分析 *代表显著性差异水平0.01<P<0.05,**代表 P <0.01,n.s.代表无显著性差异

Fig. 6 Differences in virulence test of different treatment groups * indicates a significant difference level 0.01<P<0.05,** indicates P <0.01,and n.s. indicates no significant difference

图7 相容性测定 A：SJ5进行涂板,分别将SJ17和SJ19点涂在平板上;B：SJ17进行涂板,分别将SJ5和SJ19点涂在平板上

Fig. 7 Compatible test results A：SJ5 is used for coating,and SJ17 and SJ19 are dot-coated on the plate.B：SJ17 is used for coating the plate,and SJ5 and SJ19 are dot-coated on the plate respectively

图8 不同处理对番茄根系的影响（45 d）红色箭头指向处代表番茄根部产生根结,即根结线虫寄生形成的瘤状突起

Fig. 8 Effects of different treatments on tomato root system（45 d） The points where the red arrows in the picture point to represents root knots on tomato roots,that is,nodules formed by root-knot nematode parasites

图9 不同处理对番茄根结线虫病的防治效果（45 d）不同小写字母表示差异显著（P < 0.05）

Fig. 9 Control effects of different treatments on tomato root-knot nematode disease（45 d） Different letters refer to significant differences（P < 0.05）

图10 不同处理对番茄植株长势的影响（45 d）

Fig.10 Agronomic traits of tomato plants under different treatments（45 d）

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