5株生防真菌对孢囊线虫的杀线活性测定

doi:10.13560/j.cnki.biotech.bull.1985.2022-0691

摘要/Abstract

摘要：

为了丰富防治线虫的真菌资源，测定了实验室分离保存的5株生防真菌粉红粘帚霉JGZ、淡紫紫孢菌Y2和BL2、哈茨木霉菌L4和球孢白僵菌BJ对甜菜孢囊线虫、大豆孢囊线虫和禾谷孢囊线虫的杀线虫活性。结果表明，淡紫紫孢菌Y2和哈茨木霉菌L4的孢囊寄生率为100%，随着作用时间增长，孢囊壁消解破裂。5株真菌发酵液随着稀释倍数的增加，线虫的校正死亡率降低。哈茨木霉菌L4的5倍发酵稀释液对甜菜孢囊线虫、大豆孢囊线虫和禾谷孢囊线虫的校正死亡率分别为96.75%、95.95%和96.35%；淡紫紫孢菌Y2的5倍发酵稀释液对甜菜孢囊线虫、大豆孢囊线虫和禾谷孢囊线虫的校正死亡率分别为97.37%、95.74%和97.77%，两者差异不显著。盆栽试验结果表明，哈茨木霉菌L4和淡紫紫孢菌Y2发酵液作用大豆孢囊线虫30 d，孢囊减退率分别为43.67%和41.87%，根长和茎粗显著高于其他处理，哈茨木霉菌L4处理地上鲜重和地下鲜重显著高于其他处理。结合孢囊寄生率、发酵液杀线活性、孢囊减退率和促生作用综合评价，哈茨木霉菌L4生防效果最好。

关键词: 粉红粘帚霉, 淡紫紫孢菌, 哈茨木霉菌, 球孢白僵菌, 孢囊线虫, 杀线活性

Abstract:

In order to enrich the nematicidal fungi resources, the nematicidal activities of Gliocladium roseum JGZ, Purpureocillum lilacinum Y2 and BL2, Trichoderma harzianum L4, Beauveria bassiana BJ against cyst nematodes of Heterodera schachtii（SBCN）, Heterodera glycines（SCN）, and Heterodera avenae（CCN）were determined in the laboratory. The results showed that the parasitic rates of P. lilacinum Y2, and T. harzianum L4 to cysts were 100%. As the action time increasing, the cyst cuticles were digested and broken. With the increase of dilution rate, the inhibition activities of five strains of fungal fermentations to cyst nematodes decreased. The corrected mortality rates of 5× fermentation broth of T. harzianum L4 to SBCN, SCN, CCN were 96.75%, 95.95% and 96.35%, respectively. The corrected mortality rates of 5× fermentation broth of P. lilacinum Y2 to SBCN, SCN, CCN were 97.37%,95.74% and 97.77% respectively. There were no significant difference between T. harzianum L4 and P. lilacinum Y2. The pot experiment results showed that cyst number treated with the fermentation broth of T. harzianum L4 and P. lilacinum Y2 on SCN for 30 d reduced by 43.67% and 41.87%, respectively. The root length and stem diameter were significantly higher than those by other treatments. The above-ground and under-ground fresh weight treated by T. harzianum L4 was higer than those by other treatments significantly. The biological control effect of T. harzianum L4 was the best, based on the comprehensive evaluation of cyst parasitism rate, nematicidal activity of fermentation broth, the reduction rate of cyst and growth promotion.

Key words: Gliocladium roseum, Purpureocillium lilacinum, Trichoderma harzianum, Beauveria bassiana, cyst nematodes, nematicidal activity

武玉环, 彭焕, 葛逢勇, 彭德良, 刘大群, 李亚宁. 5株生防真菌对孢囊线虫的杀线活性测定[J]. 生物技术通报, 2022, 38(11): 220-226.

WU Yu-huan, PENG Huan, GE Feng-yong, PENG De-liang, LIU Da-qun, LI Ya-ning. Nematicidal Activities Determination of Five Biocontrol Fungi Against Cyst Nematodes[J]. Biotechnology Bulletin, 2022, 38(11): 220-226.

图/表 6

参考文献 33

[1]	Folkertsma RT. Inter- and intraspecific variation between populations of Globodera rostochiensis and G. Pallida revealed by random amplified polymorphic DNA[J]. Phytopathology, 1994, 84(8): 807. doi: 10.1094/Phyto-84-807 URL
[2]	Kort J, Ross H, Stone AR, et al. An international scheme for identifying and classifying pathotypes of potato cyst-Nematodes Globodera rostochiensis and G. pallida[J]. Nematologica, 1977, 23(3): 333-339. doi: 10.1163/187529277X00057 URL
[3]	Ramsay G, Bryan G, Castelli L, et al. Investigation of resistance specificity amongst fifteen wild Solanum species to a range of Globodera pallida and G. rostochiensis populations[J]. Nematology, 2005, 7(5): 689-699. doi: 10.1163/156854105775143017 URL
[4]	Fullaondo A, Barrena E, Viribay M, et al. Identification of potato cyst nematode species Globodera rostochiensis and G. pallida by PCR using specific primer combinations[J]. Nematology, 1999, 1(2): 157-163. doi: 10.1163/156854199508126 URL
[5]	Būda V, Cepulytė-Rakauskienė R. The effect of linalool on second-stage juveniles of the potato cyst nematodes Globodera rostochiensis and G. pallida[J]. J Nematol, 2011, 43(3/4): 149-151.
[6]	Byrne JT, Maher NJ, Jones PW. Comparative responses of Globodera rostochiensis and G. pallida to hatching chemicals[J]. J Nematol, 2001, 33(4): 195-202. pmid: 19265881
[7]	Cui L, Sun L, Gao X, et al. The impact of resistant and susceptible wheat cultivars on the multiplication ofHeterodera filipjevi and H. avenae in parasite-infested soil[J]. Plant Pathol, 2016, 65(7): 1192-1199. doi: 10.1111/ppa.12495 URL
[8]	Luo SJ, Kong LG, Peng H, et al. Golden Promise barley(Hordeum vulgare)is a suitable candidate model host for investigation interaction with Heterodera avenae[J]. J Integr Agric, 2017, 16(7): 1537-1546. doi: 10.1016/S2095-3119(16)61595-2 URL
[9]	Baklawa M, Niere B, Heuer H, et al. Characterisation of cereal cyst nematodes in Egypt based on morphometrics, RFLP and rDNA-ITS sequence analyses[J]. Nematology, 2015, 17(1): 103-115. doi: 10.1163/15685411-00002855 URL
[10]	Yavuzaslanoglu E, Elekcioglu HI, Nicol JM, et al. Distribution, frequency and occurrence of cereal nematodes on the Central Anatolian Plateau in Turkey and their relationship with soil physicochemical properties[J]. Nematology, 2012, 14(7): 839-854. doi: 10.1163/156854112X631926 URL
[11]	Nour SM, Lawrence JR, Zhu H, et al. Bacteria associated with cysts of the soybean cyst nematode(Heterodera glycines)[J]. Appl Environ Microbiol, 2003, 69(1): 607-615. doi: 10.1128/AEM.69.1.607-615.2003 URL
[12]	Lian Y, Guo JQ, Li HC, et al. A new race(X12)of soybean cyst nematode in China[J]. J Nematol, 2017, 49(3): 321-326. pmid: 29062156
[13]	Meyer S, Huettel R, Liu XZ, et al. Activity of fungal culture filtrates against soybean cyst nematode and root-knot nematode egg hatch and juvenile motility[J]. Nematology, 2004, 6(1): 23-32. doi: 10.1163/156854104323072883 URL
[14]	Ali MA, Wieczorek K, Kreil DP, et al. The beet cyst nematode Heterodera schachtii modulates the expression of WRKY transcription factors in syncytia to favour its development in Arabidopsis roots[J]. PLoS One, 2014, 9(7): e102360.
[15]	Davies LJ, Lilley CJ, Paul Knox J, et al. Syncytia formed by adult female Heterodera schachtii in Arabidopsis thaliana roots have a distinct cell wall molecular architecture[J]. New Phytol, 2012, 196(1): 238-246. doi: 10.1111/j.1469-8137.2012.04238.x pmid: 22803660
[16]	Griffin GD. Differences in the response of certain weed host populations to Heterodera schachtii[J]. J Nematol, 1982, 14(2): 174-182. pmid: 19295693
[17]	Hewezi T, Piya S, Qi MS, et al. Arabidopsis miR827 mediates post-transcriptional gene silencing of its ubiquitin E3 ligase target gene in the syncytium of the cyst nematode Heterodera schachtii to enhance susceptibility[J]. Plant J, 2016, 88(2): 179-192. doi: 10.1111/tpj.13238 URL
[18]	彭德良, Subbotin S, Moens M. 小麦禾谷胞囊线虫(Heterodera avenae)的核糖体基因(rDNA)限制性片段长度多态性研究[J]. 植物病理学报, 2003, 33(4): 323-329.
	Peng DL, Subbotin S, Moens M. rDNA restriction fragment length polymorphism of Heterodera avenae in China[J]. Acta Phytopathol Sin, 2003, 33(4): 323-329.
[19]	赵洪海, 丁海燕. 2012—2013年山东省小麦禾谷孢囊线虫发生分布调查[J]. 山东农业科学, 2014, 46(4): 83-86, 91.
	Zhao HH, Ding HY. Investigation on occurrence and distribution of cereal cyst nematode in wheat fields of Shandong Province in 2012-2013[J]. Shandong Agric Sci, 2014, 46(4): 83-86, 91.
[20]	苏致衡, 黄文坤, 郑国栋, 等. 北京地区小麦禾谷孢囊线虫病发生动态调查[J]. 植物保护, 2013, 39(1): 116-120.
	Su ZH, Huang WK, Zheng GD, et al. Investigation on the occurrence of Heterodera avenae in Beijing[J]. Plant Prot, 2013, 39(1): 116-120.
[21]	彭德良. 植物线虫病害:我国粮食安全面临的重大挑战[J]. 生物技术通报, 2021, 37(7): 1-2.
	Peng DL. Plant nematode diseases: serious challenges to China’s food security[J]. Biotechnol Bull, 2021, 37(7): 1-2.
[22]	乔精松, 彭德良, 刘慧, 等. 甜菜孢囊线虫在我国的寄主范围及生活史研究[J]. 植物保护, 2021, 47(3): 177-183.
	Qiao JS, Peng DL, Liu H, et al. The host range and life history of sugarbeet cyst nematode Heterodera schachtii in China[J]. Plant Prot, 2021, 47(3): 177-183.
[23]	Peng H, Liu H, Gao L, et al. Identification of Heterodera schachtii on sugar beet in Xinjiang uygur autonomous region of China[J]. J Integr Agric, 2022, 21(6): 1694-1702. doi: 10.1016/S2095-3119(21)63797-8 URL
[24]	Xiang N, Lawrence KS, Kloepper JW, et al. Biological control of Heterodera glycines by spore-forming plant growth-promoting rhizobacteria(PGPR)on soybean[J]. PLoS One, 2017, 12(7): e0181201.
[25]	屈楠. 大豆孢囊线虫生防细菌的分离、鉴定及应用基础研究[D]. 杭州: 浙江大学, 2019.
	Qu N. Isolation, identification and application of bacterial biocontrol agents to Heterodera glycines[D]. Hangzhou: Zhejiang University, 2019.
[26]	陈井生, 陈立杰, 刘大伟, 等. 放线菌Snea49的种类鉴定及对胞囊线虫的活性评价[J]. 大豆科学, 2010, 29(4): 663-665.
	Chen JS, Chen LJ, Liu DW, et al. Indentification of the strain Snea49 and evaluation for nematicidal activity against soybean cyst nematode[J]. Soybean Sci, 2010, 29(4): 663-665.
[27]	李婷, 黄文坤, 彭德良, 等. 3株生防真菌发酵液对大豆孢囊线虫的防治效果[J]. 华中农业大学学报, 2017, 36(1): 42-46.
	Li T, Huang WK, Peng DL, et al. Control efficiency of three fungal strains’ fermentation broth on soybean cyst nematode(Heterodera glycines)[J]. J Huazhong Agric Univ, 2017, 36(1): 42-46.
[28]	张辉民, 黄文坤, 孔令安, 等. 禾谷孢囊线虫生防真菌的分离鉴定及初步应用[J]. 华北农学报, 2013, 28(4): 190-194. doi: 10.3969/j.issn.1000-7091.2013.04.035
	Zhang HM, Huang WK, Kong LG, et al. Identification and preliminary application of biocontrol fungi from cereal cyst nematode[J]. Acta Agric Boreali Sin, 2013, 28(4): 190-194.
[29]	陈秀菊, 李惠霞, 徐志鹏, 等. 3株生防真菌的杀线虫活性及种类鉴定[J]. 大豆科学, 2019, 38(4): 576-583.
	Chen XJ, Li HX, Xu ZP, et al. Nematocidal activities and species identification of three biocontrol fungi[J]. Soybean Sci, 2019, 38(4): 576-583.
[30]	田忠玲. 大豆孢囊线虫生防真菌的筛选、鉴定及应用基础研究[D]. 杭州: 浙江大学, 2016.
	Tian ZL. Isolation, identification and application of fungal biocontrol agents to Heterodera glycines[D]. Hangzhou: Zhejiang University, 2016.
[31]	Jin N, Liu SM, Peng H, et al. Isolation and characterization of Aspergillus niger NBC001 underlying suppression against Heterodera glycines[J]. Sci Rep, 2019, 9(1): 591. doi: 10.1038/s41598-018-37827-6 pmid: 30679719
[32]	陈倩, 张露源, 陈伯昌, 等. 大豆孢囊线虫生防菌株Myrothecium verrucaria ZW-2发酵条件优化及活性物质分析[J]. 生物技术通报, 2021, 37(7): 127-136. doi: 10.13560/j.cnki.biotech.bull.1985.2021-0454
	Chen Q, Zhang LY, Chen BC, et al. Optimization of fermentation conditions of Myrothecium verrucaria ZW-2, a biocontrol strain against Heterodera glycines and analysis of active substances[J]. Biotechnol Bull, 2021, 37(7): 127-136.
[33]	张春龙, 肖炎农, 向妮, 等. 淡紫拟青霉防治小麦禾谷孢囊线虫病研究[J]. 植物保护, 2014, 40(4): 181-184.
	Zhang CL, Xiao YN, Xiang N, et al. Biological control effect of Paecilomyces lilacinus against cereal cyst nematode(CCN)[J]. Plant Prot, 2014, 40(4): 181-184.

菌株Strain	稀释倍数 Dilution rate	甜菜孢囊线虫 Heterodera schachtii	大豆孢囊线虫 Heterodera glycines	禾谷孢囊线虫 Heterodera avenae
粉红粘帚霉（G. roseum）JGZ	5×	85.58±3.21Ac	86.61±2.92Ab	98.18±0.86Aa
	10×	78.51±4.66Bc	83.99±3.68Ab	87.85±2.97Bb
	15×	75.86±2.80Bc	82.37±3.77ABb	75.70±4.32Cd
	20×	64.04±4.01Cc	77.94±4.23Bc	71.72±2.94Cc
淡紫紫孢菌（P. lilacinum）BL2	5×	95.95±1.59Aa	89.67±4.47Ab	96.56±2.09Aa
	10×	95.73±2.55Aa	82.96±3.15Bb	88.47±2.70Bb
	15×	80.72±2.64Bb	61.27±3.25Cd	80.16±1.13Cbc
	20×	75.96±3.38Cb	53.64±4.53Dd	74.95±5.17Dc
哈茨木霉菌（T. harzianum）L4	5×	96.75±1.32Aa	95.95±2.57Aa	96.35±1.36Aa
	10×	94.53±3.48Aa	93.91±3.14ABa	87.44±2.02Bb
	15×	83.77±2.71Bb	93.51±1.35ABa	76.73±3.15Ccd
	20×	78.39±2.21Cab	92.31±2.19Ba	60.81±3.53Dd
淡紫紫孢菌（P. lilacinum）Y2	5×	97.37±1.35Aa	95.74±1.66Aa	97.77±0.84Aa
	10×	95.73±2.22Aa	92.49±2.25ABa	93.52±1.36Ba
	15×	94.32±3.00Aa	88.65±3.98BCa	90.69±1.64Ca
	20×	84.24±3.08Ba	84.41±4.33Cb	86.67±2.41Da
球孢白僵菌（B. bassiana）BJ	5×	92.09±2.82Ab	82.35±3.01Ac	94.13±2.53Ab
	10×	84.97±5.10Ab	77.68±3.67Ac	85.02±5.01Bb
	15×	70.00±4.79Bd	68.79±6.49Bc	82.59±3.03Bb
	20×	59.19±9.85Cc	34.41±3.43Ce	82.02±2.80Bb

菌株Strain	稀释倍数 Dilution rate	甜菜孢囊线虫 Heterodera schachtii	大豆孢囊线虫 Heterodera glycines	禾谷孢囊线虫 Heterodera avenae
粉红粘帚霉（G. roseum）JGZ	5×	85.58±3.21Ac	86.61±2.92Ab	98.18±0.86Aa
	10×	78.51±4.66Bc	83.99±3.68Ab	87.85±2.97Bb
	15×	75.86±2.80Bc	82.37±3.77ABb	75.70±4.32Cd
	20×	64.04±4.01Cc	77.94±4.23Bc	71.72±2.94Cc
淡紫紫孢菌（P. lilacinum）BL2	5×	95.95±1.59Aa	89.67±4.47Ab	96.56±2.09Aa
	10×	95.73±2.55Aa	82.96±3.15Bb	88.47±2.70Bb
	15×	80.72±2.64Bb	61.27±3.25Cd	80.16±1.13Cbc
	20×	75.96±3.38Cb	53.64±4.53Dd	74.95±5.17Dc
哈茨木霉菌（T. harzianum）L4	5×	96.75±1.32Aa	95.95±2.57Aa	96.35±1.36Aa
	10×	94.53±3.48Aa	93.91±3.14ABa	87.44±2.02Bb
	15×	83.77±2.71Bb	93.51±1.35ABa	76.73±3.15Ccd
	20×	78.39±2.21Cab	92.31±2.19Ba	60.81±3.53Dd
淡紫紫孢菌（P. lilacinum）Y2	5×	97.37±1.35Aa	95.74±1.66Aa	97.77±0.84Aa
	10×	95.73±2.22Aa	92.49±2.25ABa	93.52±1.36Ba
	15×	94.32±3.00Aa	88.65±3.98BCa	90.69±1.64Ca
	20×	84.24±3.08Ba	84.41±4.33Cb	86.67±2.41Da
球孢白僵菌（B. bassiana）BJ	5×	92.09±2.82Ab	82.35±3.01Ac	94.13±2.53Ab
	10×	84.97±5.10Ab	77.68±3.67Ac	85.02±5.01Bb
	15×	70.00±4.79Bd	68.79±6.49Bc	82.59±3.03Bb
	20×	59.19±9.85Cc	34.41±3.43Ce	82.02±2.80Bb

处理 Treatment	株高 Height/cm	根长 Length of root/cm	茎粗 Stem diameter/cm	地上鲜重 Above-ground fresh weight/g	地下鲜重 Under-ground fresh weight/g
粉红粘帚霉（G. roseum）JGZ	36.56±2.20a	14.56±1.36b	0.31±0.01b	1.90±0.17c	0.22±0.07b
淡紫紫孢菌（P. lilacinum）BL2	34.64±1.87a	13.52±1.09b	0.32±0.02b	2.15±0.34bc	0.23±0.05b
哈茨木霉菌（T. harzianum）L4	35.36±1.13a	20.30±1.12a	0.35±0.03a	2.83±0.13a	0.39±0.04a
淡紫紫孢菌（P. lilacinum）Y2	36.04±1.50a	18.88±0.86a	0.35±0.03a	2.31±0.15b	0.29±0.04b
球孢白僵菌（B. bassiana）BJ	36.20±1.28a	14.35±1.47b	0.31±0.01b	1.83±0.10c	0.24±0.07b
CK	35.37±0.57a	13.87±1.50b	0.32±0.01b	2.10±0.34bc	0.24±0.04b

处理 Treatment	株高 Height/cm	根长 Length of root/cm	茎粗 Stem diameter/cm	地上鲜重 Above-ground fresh weight/g	地下鲜重 Under-ground fresh weight/g
粉红粘帚霉（G. roseum）JGZ	36.56±2.20a	14.56±1.36b	0.31±0.01b	1.90±0.17c	0.22±0.07b
淡紫紫孢菌（P. lilacinum）BL2	34.64±1.87a	13.52±1.09b	0.32±0.02b	2.15±0.34bc	0.23±0.05b
哈茨木霉菌（T. harzianum）L4	35.36±1.13a	20.30±1.12a	0.35±0.03a	2.83±0.13a	0.39±0.04a
淡紫紫孢菌（P. lilacinum）Y2	36.04±1.50a	18.88±0.86a	0.35±0.03a	2.31±0.15b	0.29±0.04b
球孢白僵菌（B. bassiana）BJ	36.20±1.28a	14.35±1.47b	0.31±0.01b	1.83±0.10c	0.24±0.07b
CK	35.37±0.57a	13.87±1.50b	0.32±0.01b	2.10±0.34bc	0.24±0.04b