Abstract:Morel (Morchella spp.) is a world-famous rare edible and medicinal mushroom. In the artificial cultivation process morels must be covered with soil to produce fruit. At present, the mechanism of soil microorganisms affecting the growth and development of morel is still unknown. 132 strains of bacteria were isolated and purified from morel soil. Through the plate confrontation culture test, it was found that 15 strains had an antagonistic effect on the hyphae of Morchella importuna and 4 strains had the effect of promoting growth. Among them, when strain X14 was co-cultured with morel on plate for 4 and 8 d, the growth rate of morel hyphae was significantly increased by 7.6% and 11.6% compared with the control (P<0.05). Morel and strain X14 were inoculated into wheat medium in test tubes and co-cultured. The growth rate of morel hyphae was measured at different culture time. The results showed that the growth rate of morel hyphae treated by strain X14 was very extremely significantly increased by 28.3%, 24.8% and 24.0% compared with the control at 4, 6 and 7 d, respectively (P<0.001). Morel and strain X14 were inoculated into stock flasks and co-cultured. The results showed that the growth rate of morel hyphae treated by strain X14 was not significantly different from the control at 5 d of co-culture (P>0.05). After 10 d of co-cultivation, the growth rate of morel hyphae treated with strain X14 was extremely significantly increased by 14.1% compared with the control (P<0.01). The culture period of the stock seed was shortened by 4~6 d compared with the control. Based on the comprehensive analysis of colony morphology, physiological and biochemical characteristics and 16S rDNA sequence, the strain X14 was identified as Pseudomonas putida. In this study, a strain of Pseudomonas putida X14 was screened from the soil of morel, which could promote the hyphae growth of Morchella importuna and provide technical support for the development of microbial agents that promote the growth of morel.
[1] 董晓雅, 周巍巍, 张继英, 等. 2010. 荧光假单胞菌对食用菌的促生作用及其机理[J]. 生态学报, 30(17): 4685-4690. (Dong X Y, Zhou W W, Zhang J Y, et al.2010. The effect of growth promotion and its mechanism on edible fungi by Pseudomonas fluorescens[J]. Journal of Ecology, 30(17): 4685-4690.) [2] 杜习慧. 2019. 黑色羊肚菌支系的物种资源、生殖模式和遗传多样性研究进展[J]. 菌物研究, 17(4): 240-251. (Du X H.2019. Review on species resources, reproductive modes and genetic diversity of black morels[J]. Journal of Fungal Research, 17(4): 240-251.) [3] 何畅, 徐丽婧, 常明昌, 等. 2021. 梯棱羊肚菌子实体多糖提取优化及结构初探[J]. 食用菌学报, 28(2): 77-88. (He C, Xu L J, Chang M C, et al.2021. Extraction optimization and structural characterization of polysaccharides from Morchella importuna fruiting body[J]. Acta Edulis Fungi, 28(2): 77-88.) [4] 何佳, 赵启美. 2000. 硅酸盐细菌G10菌株对食用菌的影响[J]. 中国食用菌, 19(3): 35-36. (He J, Zhao Q M.2000. Effects of silicate bacteria G10 strain on edible mushrooms[J]. Edible Fungi of China, 19(3): 35-36.) [5] 阚洪媛, 杨世鑫, 孙梁伦, 等. 2020. 一株耐铅、锌、铬菌株的分离鉴定及其吸附能力[J]. 微生物学通报, 47(12): 3974-3986. (Kan H Y, Yang S X, Sun L L, et al.2020. Isolation, identification and adsorption capacity of a strain resistant to lead, zinc and chromium[J]. Microbiology China, 47(12): 3974-3986.) [6] 林彬, 赵学才, 刘宜敏, 等. 1997. 河南新郑地区野生羊肚菌属的种类及生态研究[J]. 河南科学, (2): 191-195. (Lin B, Zhao X C, Liu Y M, et al. 1997. Investigation on the kinds of wild morchella and their ecological studies at Xin Zheng district[J]. Henan Science, (2): 191-195.) [7] 刘伟, 张亚, 何培新. 2017. 羊肚菌生物学与栽培技术[M]. 长春: 吉林科学技术出版社, pp. 292-302. (Liu W, Zhang Y, He P X.2017. Biology and cultivation technique of morchella[M]. Jilin Science and Technology Press, Changchun, China, pp. 292-302.) [8] 刘娜, 宋莹, 吕立涛, 等. 2019. 5株梯棱羊肚菌菌丝生长特性研究[J]. 中国食用菌. 38(9): 21-28. (Liu N, Song Y, Lv L T, et al.2019. Study on mycelia growth characters of 5 Morchella importuna strains[J]. Edible Fungi of China, 38(9): 21-28.) [9] 刘奇正, 董彩虹. 2020. 羊肚菌菌核的形成研究进展及其在栽培中应用的探讨[J]. 食用菌学报, 27(4): 172-178. (Liu Q Z, Dong C H.2020. Research progress on sclerotium formation in morchella and reflection on its application in cultivation[J]. Acta Edulis Fungi, 27(4): 172-178.) [10] 谭方河. 2016. 羊肚菌人工栽培技术的历史、现状及前景[J]. 食药用菌, 24(3): 140-144. (Tan F H.2016. History, present situation and prospect of artificial cultivation technique of morchella[J]. Edible and Medicinal Mushrooms, 24(3): 140-144.) [11] 王琳, 魏启舜, 周影, 等. 2018. 覆土层益生菌恶臭假单胞菌TK3对双孢蘑菇的促生作用[J]. 食用菌学报, 25(3): 23-29. (Wang L, Wei Q S, Zhou Y, et al.2018. Addition of Pseudomonas putida TK3 into the casing soil to promote growth of Agaricus bisporus[J]. Acta Edulis Fungi, 25(3): 23-29.) [12] 王龙, 周庆平, 刘建明. 2020. 基于rDNA-ITS序列的甘肃甘南野生羊肚菌遗传多样性分析[J]. 江苏农业科学, 48(17): 81-84. (Wang L, Zhou Q P, Liu J M.2020. Genetic diversity analysis of wild morchella in Gannan, Gansu based on rDNA-ITS sequence[J]. Jiangsu Agricultural Science, 48(17): 81-84.) [13] 熊川, 李小林, 李强, 等. 2015. 羊肚菌菌塘土壤细菌群落的结构及多样性[J]. 湖南农业大学学报(自然科学版), 41(4): 428-434. (Xiong C, Li X L, Li Q, et al.2015. Bacteria community structure and diversity in morchella colonies[J]. Journal of Hunan Agricultural University (Natural Sciences), 41(4): 428-434.) [14] 杨晓绒, 赖晓辉, 吾尔恩•阿合别尔迪, 等. 2020. 昭苏县野生羊肚菌根际土壤细菌多样性研究[J]. 微生物学杂志, 40(4): 24-33. (Yang X R, Lai X H, Oren A, et al.2020. Bacterial diversity in rhizosphere soil of wild morels in Zhaosu County[J]. Journal of Microbiology, 40(4): 24-33.) [15] 张相锋, 杨晓绒, 焦子伟. 2019. 伊犁野生羊肚菌根际土壤微生物功能多样性分析[J]. 微生物学杂志, 39(2): 83-89. (Zhang X F, Yang X R, Jiao Z W.2019. Functional diversity analysis of rhizosphere soil microorganism of wild morchella in Yily[J]. Journal of Microbiology, 39(2): 83-89.) [16] Benucci G M, Longley R, Zhang Peng, et al.2019. Microbial communities associated with the black morel Morchella sextelata cultivated in greenhouses[J]. Peer Journal, 7: e7744. [17] Colauto N B, Fermor, T R, Eira A F, et al.2016. Pseudomonas putida stimulates primordia on Agaricus bitorquis[J]. Current Microbiology, 72(4): 482-488. [18] Hassan W, Bano R, Bashir F,et al.2014. Comparative effectiveness of ACC-deaminase and/or nitrogen-fixing rhizobacteria in promotion of maize (Zea mays L.) growth under lead pollution[J]. Environmental Science and Pollution Research International, 21(18): 10983-10996. [19] Kim M K, Math P K, Cho K M, et al.2008. Effect of Pseudomonas sp. P7014 on the growth of edible mushroom Pleurotus eryngii in the bottle culture for commercial production[J]. Bioresource Technology, 99(8): 3306-3308. [20] Penrose D M, Glick B R.2003. Methods for isolating and characterizing ACC deaminase-containing plant growth-promoting rhizobacteria[J]. Physiology Plant, 118(1): 10-15. [21] Pion M, Spangenberg J E, Simon A, et al.2013. Bacterial farming by the fungus Morchella crassipes[J]. Proceedings of Royal Society B-Biological Sciences, 280: 20132242. [22] Yu F M, Jayawardena R S, Thongklang N, et al.2022. Morel production associated with soil nitrogen-fixing and nitrifying microorganisms[J]. Journal of Fungi, 8(3): 299.