哈茨木霉菌对烟草的促生及其黑胫病的诱导抗性评价

doi:10.13304/j.nykjdb.2024.0225

摘要/Abstract

摘要：

为研究哈茨木霉菌（Trichoderma harzianum）对烟草促生作用及由烟草疫霉菌（Phytophthora nicotianae）引起的烟草黑胫病的诱导抗性分子机制，对5株木霉菌株进行筛选。结果表明，哈茨木霉ACCC 32808（TH32808）菌株对烟草疫霉菌的抑制性较强，抑制率为62.00%。进一步研究TH32808对不同烟草品种的促生作用及对烟草黑胫病的生物防治诱导抗性，结果表明，在盆栽条件下，TH32808对不同烟草品种的促生作用不同，其中红花大金元的植株鲜重显著增加8.30%，对黑胫病害的防治效果可达51.74%；红花大金元和K326的过氧化氢酶活性在第4天时较高，分别为对照的1.42和2.15倍，过氧化物酶活性在第2天时较高，分别为对照的2.49和2.29倍，超氧化物歧化酶活性在第4天较高，分别为对照的1.17和1.15倍。TH32808可诱导烟草对黑胫病的抗性，关键信号分子水杨酸、茉莉酸和乙烯代谢途径的关键基因NPR1、PDF1.2、EFE和NtSGT1均呈上调表达趋势。研究结果为烟草黑胫病的生物防治和诱导抗性研究提供了生防真菌资源和理论依据。

关键词: 哈茨木霉, 促生作用, 烟草黑胫病, 诱导抗性, 生物防治

Abstract:

To investigate the growth-promoting effects of Trichoderma harzianum on tobacco and its induced resistance mechanisms against tobacco black shank disease caused by Phytophthora nicotianae， 5 Trichoderma strains were screened. The results showed that the strain T. harzianum ACCC 32808 （TH32808） was demonstrated the strongest inhibition （62.00%） among 5 strains of Trichoderma. The growth-promoting effects of TH32808 on different tobacco varieties and its induced resistance against tobacco black shank disease were assessed. The results showed that， under pot condition， TH32808 exhibited different ability on promoting the growth of different tobacco varieties. Under TH32808 treatment， the fresh weight of Honghuadajinyuan increased by 8.30%， the control efficacy against black shank disease in Honghuadajinyuan was 51.74%. In Honghuadajinyuan and K326， the catalase activity peaked on the 4th day， reaching 1.42 and 2.15 times that of CK， the peroxidase activity peaked on the 2nd day， being 2.49 and 2.29 times that of CK， the superoxide dismutase activity peaked on the 4th day， reaching 1.17 and 1.15 times that of CK， respectively. Moreover， TH32808 could induce resistance against black shank disease， key genes involved in the metabolic pathways of salicylic acid， jasmonic acid and ethylene， such as NPR1， PDF1.2， EFE and NtSGT1， all exhibited an upregulation trend. Above results provided fungal resources and theoretical basis for the biological control and induced resistance research against black shank disease in tobacco.

Key words: Trichoderma harzianum, growth-promoting effect, black shank disease in tobacco, induced resistance, biological control

中图分类号:

S572

常峻嘉, 盖佳鑫, 陶刚, 莫转龙海. 哈茨木霉菌对烟草的促生及其黑胫病的诱导抗性评价[J]. 中国农业科技导报, 2024, 26(10): 168-176.

Junjia CHANG, Jiaxin GAI, Gang TAO, Zhuanlonghai MO. Evaluation of the Growth-promoting Effect of Trichoderma harzianum on Tobacco and Its Induced Resistance to Black Shank Disease[J]. Journal of Agricultural Science and Technology, 2024, 26(10): 168-176.

图/表 9

表1 实时定量PCR分析的基因和引物

Table 1 Genes and primers for real-time qPCR analysis

基因 Gene	上游引物 Forward primer （5’-3’）	下游引物 Reverse primer（5’-3’）
β-actin	ATGCCTATGTGGGTGACGAAG	TCTGTTGGCCTTAGGGTTGAG
NPR1	ACATCAGCGGAAGCAGTAG	GTCGGCGAAGTAGTCAAAC
NtPDF1.2	GGAAATGGCAAACTCCATGCG	ATCCTTCGGTCAGACAAACG
EFE	ATTTATCCAGCACCAGCTCT	GCTTCAAACCTTGGCTCCT
NtSGT1	TCCACACTCGCCATCCTGAA	GTGCCGCAAATTCAACTCTG

图1 不同木霉对烟草疫霉菌的抑制作用A：TH 32808；B：ACCC 32809；C：ACCC 31649；D：ACCC 32826；E：ACCC 32969；F：CK1

Fig. 1 Inhibition of different kinds of Trichoderma on Phytophthora nicotianae

表2 不同木霉处理对烟草疫霉菌的抑制率

Table 2 Inhibition ratio of different kinds of Trichoderma treatments on Phytophthora nicotianae

处理 Treatment	病原菌半径 Pathogen radius/mm	抑制率 Inhibition rate/%
TH 32808	7.17±0.21 d	62.00
ACCC 32809	7.84±0.33 bcd	58.45
ACCC 31649	8.49±0.65 bc	54.96
ACCC 32826	7.36±0.16 cd	60.07
ACCC 32969	8.83±0.43 b	53.16
CK1	18.86±0.44 a	—

表3 TH32808和TH31649对烟草植株的促生作用

Table 3 Growth promotion effect of TH32808 and TH31649 on tobacco plants

品种 Variety	处理 Treatment	株高 Plant height/cm	茎径 Stem diameter/mm	鲜重 Fresh weight/g	干重 Dry weight/g
红花大金元 Honghuadajinyuan	CK2	75.58±6.54 a	5.05±0.11 ab	12.99±0.37 b	1.20±0.04 b
	TH32808	72.36±4.67 a	5.23±0.24 a	13.93±0.68 ab	1.16±0.06 b
	ACCC31649	66.19±6.32 ab	4.85±0.15 ab	14.05±0.46 a	1.52±0.04 a
K326	CK2	49.82±2.47 bc	4.46±0.19 b	9.47±0.42 c	0.76±0.44 cd
	TH32808	40.86±3.71 c	4.83±0.31 ab	9.59±0.41 c	0.74±0.02 d
	ACCC31649	45.23±4.61 c	4.52±0.11 b	10.25±0.56 c	0.83±0.05 c

表4 TH32808对红花大金元烟草黑胫病的盆栽防治效果

Table 4 Effect of TH32808 on the black shank disease of the potted Honghuadajinyuan plants

处理

Treatment

发病率

Incidence rate/%

病情指数

Disease index

防治效果

Control effect/%

图2 TH32808处理后对红花大金元和K326的酶活性注：不同小写字母表示同一时间不同处理间差异在P<0.05水平差异显著。

Fig. 2 Enzyme activity of Honghuadajinyuan and K326 leaves under TH32808 treatmentNote： Different lowercase letters indicate significant differences between different treatments of same time at P<0.05 level.

图3 烟草疫霉菌感染下接种TH32808后红花大金元和K326的酶活力注：不同小写字母表示同一时间不同处理间在P<0.05水平差异显著。

Fig. 3 Enzyme activity of Honghuadajinyuan and K326 leavesinfected Phytophthora nicotiana with TH32808Note： Different lowercase letters indicate significant differences between different treatments of same time at P<0.05 level

图4 TH32808处理下红花大金元抗性基因的表达注：不同小写字母表示不同处理间在P<0.05水平差异显著。

Fig. 4 Expression of resistance genes in Honghuadajinyuan treated by TH32808Note： Different lowercase letters indicate significant differences between different treatments at P<0.05 level.

图5 TH32808处理下K326抗性基因的表达注：不同小写字母表示不同处理间在P<0.05水平差异显著。

Fig. 5 Expression of resistance genes in K326 treated by TH32808Note： Different lowercase letters indicate significant differences between different treatments at P<0.05 level.

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