青霉菌灭活菌丝体对白车轴草和黑麦草生长及生理特性的影响

doi:10.17521/cjpe.2024.0188

摘要/Abstract

摘要：

施用生长促进剂是人工辅助干预加快退化草地生态恢复的重要手段, 为挖掘适用于退化草地的生长促进剂, 该研究采用盆栽实验和田间实验的方法, 探索了青霉素工业生产过程中产生的残余物青霉菌灭活菌丝体(DMP)对禾豆混播法常用牧草白车轴草(Trifolium repens)和黑麦草(Lolium perenne)生长生理的影响。研究结果表明: (1) DMP能够显著改善白车轴草和黑麦草的生理指标, 并改良二者的发育形态学性状。盆栽条件下每盆施用4.50 g DMP显著增加了黑麦草的株高和叶片数, 与复合肥处理相比分别增加了27.59%和44.16%, 而每盆施用2.25 g DMP显著增加了黑麦草的分蘖数, 与复合肥处理相比增加了38.79%。同时, 每盆施用2.25 g DMP显著增加了白车轴草的分支数和叶片数, 与复合肥处理相比分别增加了27.59%和44.16%。与复合肥处理相比, 每盆4.50 g DMP处理显著提高了黑麦草的叶绿素总量和粗脂肪含量, 提高率分别为12.77%和11.82%, 而每盆6.00 g DMP显著提高了黑麦草的粗蛋白含量, 提高率为119.94%。每盆4.50 g DMP处理也显著提高了白车轴草的叶绿素总量和粗蛋白含量, 与复合肥处理相比提高率分别为29.47%和42.46%。(2) DMP能够提高黑麦草和白车轴草的生物量。盆栽条件下, 与复合肥处理相比, 每盆3.40 g DMP处理显著提高了黑麦草和白车轴草的地上部分干质量, 提高率分别为22.76%和21.09%。在昭通低温寡日照田间条件下, 与复合肥处理相比, 225 kg·hm^-2 DMP提高了黑麦草的地上部分和地下部分干质量积累, 提高率分别为64.53%和14.64%, 225 kg·hm^-2 DMP也提高了白车轴草的地上部分和地下部分干质量积累, 提高率分别为58.00%和129.19%。(3)转录组测序表明, DMP处理诱导了黑麦草植物激素生物合成和信号转导通路相关基因的表达, 证实DMP通过诱导激素生物合成和信号转导通路促进牧草生长。该研究首次证实了DMP能够促进牧草生长, 提出了使用DMP作为牧草生长促进剂的新策略, 为低温寡日照条件下退化草地的修复及可持续发展提供了新思路。

关键词: 退化草地恢复, 青霉菌灭活菌丝体, 促生长作用, 白车轴草, 黑麦草

Abstract:

Aims The application of growth promoter is an important tool for ecological restoration of degraded grassland by human intervention. Therefore, the aim of this study is to investigate the growth promoting effect of dry mycelium of Penicillium chrysogenum (DMP), a residue from the industrial production of penicillin, on the growth and physiological performance of Trifolium repensand Lolium perenne.

Methods In this study, we mainly applied pot experiment and field experiment to investigate the growth promoting effect of DMP addition on the performance of T. repens and L. perenne. Then we applied transcriptome sequencing to determine the molecular mechanism of DMP promoting effects on forage growth.

Important findings The results of this study are as follows: (1) DMP addition can significantly improve their physiological indices and enhanced developmental morphological traits of T. repens and L. perenne. Compared with the compound fertilizer treatment, the application of 4.50 g DMP per pot significantly increased the plant height, tiller number and leaf number of L. perenne, and the application of 2.25 g DMP per pot significantly increased the plant height, branching number and leaf number of T. repens, while 4.50 g·pot^-1 of DMP significantly increased the total chlorophyll content, crude protein content and crude fat content of L. perenne, and this concentration of DMP addition also significantly increased the total chlorophyll and crude protein contents of T. repens. (2) DMP addition can increase the biomass of L. perenne and T. repens. Under potting conditions, DMP application significantly increased aboveground fresh and dry mass of L. perenneand T. repens compared with compound fertilizer treatment. DMP at 225 kg·hm^-2 also increased aboveground and belowground dry mass accumulation of L. perenneand T. repensunder low temperature and low sunlight field conditions in Zhaotong. (3) Transcriptome sequencing showed that DMP treatment induced the expression of genes related to phytohormone biosynthesis and signal transduction pathways in L. perenne, confirming that DMP promotes forage growth by inducing hormone biosynthesis and signal transduction pathways. This study first demonstrates that DMP can promote the growth of forage, and proposes a new strategy of using DMP as a growth promoter, which provides new ideas for the restoration and sustainable development of degraded grasslands under low temperature and low sunlight conditions.

Key words: restoration of the degraded grassland, dry mycelium of Penicillium chrysogenum, growth promotion, Trifolium repens, Lolium perenne

夏敏菖, 李倩倩, 钱清清, 任淑君, 梁应冲, 陈亭颖, 李映佳, 牟宗敏, 陈穗云. 青霉菌灭活菌丝体对白车轴草和黑麦草生长及生理特性的影响. 植物生态学报, 2025, 49(1): 189-198. DOI: 10.17521/cjpe.2024.0188

XIA Min-Chang, LI Qian-Qian, QIAN Qing-Qing, REN Shu-Jun, LIANG Ying-Chong, CHEN Ting-Ying, LI Ying-Jia, MOU Zong-Min, CHEN Sui-Yun. Effect of dry mycelium of Penicillium chrysogenum on the growth and physiological performance of Trifolium repens and Lolium perenne. Chinese Journal of Plant Ecology, 2025, 49(1): 189-198. DOI: 10.17521/cjpe.2024.0188

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链接本文: https://www.plant-ecology.com/CN/10.17521/cjpe.2024.0188

https://www.plant-ecology.com/CN/Y2025/V49/I1/189

图/表 4

图1 青霉菌灭活菌丝体(DMP)对白车轴草和黑麦草形态指标的影响。Control, 空白对照; Fertilizer, 复合肥处理; n, 样本量。不同小写字母表示不同处理间存在显著差异(p < 0.05)。

Fig. 1 Effect of dry mycelium of Penicillium chrysogenum (DMP) on morphological indicators of T. repens and L. perenne. n, sample size. Different lowercase letters mean significant differences among different treatments (p < 0.05).

图2 青霉菌灭活菌丝体(DMP)对白车轴草和黑麦草生理指标的影响(平均值±标准差)。Control, 空白对照; Fertilizer, 复合肥处理; n, 样本量。不同小写字母表示不同处理间存在显著差异(p < 0.05)。

Fig. 2 Effect of dry mycelium of Penicillium chrysogenum (DMP) on the physiological indicators of T. repens and L. perenne (mean ± SD). n, sample size. Different lowercase letters mean significant differences among different treatments (p < 0.05).

图3 青霉菌灭活菌丝体(DMP)对白车轴草和黑麦草生物量的影响(平均值±标准差)。A-D为盆栽条件, E-H为田间条件。 Control, 空白对照; Fertilizer, 复合肥处理; n, 样本量。不同小写字母表示不同处理间存在显著差异(p < 0.05)。

Fig. 3 Effect of dry mycelium of Penicillium chrysogenum (DMP) on the biomass of T. repens and L. perenne (mean ± SD). A-D were under pot condition, E-H were under field condition. n, sample size. Different lowercase letters mean significant differences among different treatments (p < 0.05).

图4 青霉菌灭活菌丝体(DMP)调控内源植物激素生物合成和信号转导相关基因。A, 空白对照 vs DMP处理组差异基因火山图。B, 空白对照 vs DMP处理组内源植物激素生物合成和信号转导相关基因表达热图。

Fig. 4 Dry mycelium of Penicillium chrysogenum (DMP) regulates the expression of endogenous plant hormone biosynthesis and signal transduction related genes. A, Volcanic plot of differentially expressed genes in control (CK) vs DMP. B, Heat map of gene expressions related to endogenous plant hormone biosynthesis and signal transduction in CK vs DMP treatment.

参考文献 35

[1]	Chen Y, Zheng X, Zhang W, Li YC, Yan CL, Yang F, Yin ZR, Li WR, Wang JG, Chen SY (2013). Insect disease prevention efficiency and production increasing function of rice by using devitalized dry mycelium of Penicillium chrysogenum. Journal of Kunming University, 35(6), 66-68.
	[陈勇, 郑仙, 张旺, 李永川, 闫春丽, 杨发, 尹忠仁, 李文荣, 王建光, 陈穗云 (2013). 青霉菌灭活菌丝体对水稻病害的防效及其增产作用. 昆明学院学报, 35(6), 66-68.]
[2]	Cohen AC, Travaglia CN, Bottini R, Piccoli PN (2009). Participation of abscisic acid and gibberellins produced by endophytic Azospirillum in the alleviation of drought effects in maize. Botany, 87, 455-462.
[3]	Deng J (2023). Restoration technology of degraded grassland in Zhaotong. Forest Inventory and Planning, 48, 207-212.
	[邓军 (2023). 昭通市退化草地修复技术研究. 林业调查规划, 48, 207-212.]
[4]	Dong H, Cohen Y (2002a). Dry mycelium of Penicillium chrysogenum induces resistance against verticillium wilt and enhances growth of cotton plants. Phytoparasitica, 30, 147-157.
[5]	Dong H, Cohen Y (2002b). Induced resistance in cotton seedlings against fusarium wilt by dried biomass of Penicillium chrysogenum and its water extract. Phytoparasitica, 30, 77-87.
[6]	Dong HZ, Li WJ, Zhang DM, Tang W (2003). Differential expression of induced resistance by an aqueous extract of killed Penicillium chrysogenum against verticillium wilt of cotton. Crop Protection, 22, 129-134.
[7]	Dong HZ, Zhang XK, Choen Y, Zhou Y, Li WJ, Li ZH (2006). Dry mycelium of Penicillium chrysogenum protects cotton plants against wilt diseases and increases yield under field conditions. Crop Protection, 25, 324-330.
[8]	Duan JB, Yu H, Yuan K, Liao ZG, Meng XB, Jing YH, Liu GF, Chu JF, Li JY (2019). Strigolactone promotes cytokinin degradation through transcriptional activation of CYTOKININ OXIDASE/DEHYDROGENASE 9 in rice. Proceedings of the National Academy of Sciences of the United States of America, 116, 14319-14324.
[9]	Duan YM, Wang XX, Xu XY, Chen L, Wang JG, Lin XH, Yang XC, Chen SY (2011). Effect of dry mycelium (dm) Penicillium chrysogenum (pen) on flue-CURED tobacco seeding-STAGE traits. Southwest China Journal of Agricultural Sciences, 24, 48-51.
	[端永明, 王晓霞, 徐兴阳, 陈林, 王建光, 林兴红, 杨新成, 陈穗云 (2011). 青霉菌灭活菌丝体对烤烟苗期性状的影响. 西南农业学报, 24, 48-51.]
[10]	Fang ZM, Ji YY, Hu J, Guo RK, Sun SY, Wang XL (2020). Strigolactones and brassinosteroids antagonistically regulate the stability of the D53-OsBZR1 complex to determine FC1 expression in rice tillering. Molecular Plant, 13, 586-597.
[11]	Fu J, Zhang SY, Wu JH, Chen Y, Zhong Y, Zhou YF, Wang JG, Chen SY (2020). Structural characterization of a polysaccharide from dry mycelium of Penicillium chrysogenum that induces resistance to Tobacco mosaic virus in tobacco plants. International Journal of Biological Macromolecules, 156, 67-79.
[12]	Fu SF, Wei JY, Chen HW, Liu YY, Lu HY, Chou JY (2015). Indole-3-acetic acid: a widespread physiological code in interactions of fungi with other organisms. Plant Signaling & Behavior, 10, e1048052. DOI: 10.1080/15592324.2015.1048052.
[13]	Galuszka P, Frébort I, Sebela M, Sauer P, Jacobsen S, Pec P (2001). Cytokinin oxidase or dehydrogenase? Mechanism of cytokinin degradation in cereals. European Journal of Biochemistry, 268, 450-461. DOI PMID
[14]	Gotlieb D, Oka YJ, Ben-Daniel BH, Cohen Y (2003). Dry mycelium of Penicillium chrysogenum protects cucumber and tomato plants against the root-knot nematode Meloidogyne javanica. Phytoparasitica, 31, 217-225.
[15]	Hou XM, Wang CY, Gu YC, Shao CL (2016). Penimethavone A, a flavone from a gorgonian-derived fungus Penicillium chrysogenum. Natural Product Research, 30, 2274-2277.
[16]	Li Y, Jiao MT, Li YJ, Zhong Y, Li XQ, Chen ZZ, Chen SY, Wang JG (2021). Penicillium chrysogenum polypeptide extract protects tobacco plants from tobacco mosaic virus infection through modulation of ABA biosynthesis and callose priming. Journal of Experimental Botany, 72, 3526-3539.
[17]	Liu KF, Li T, Duan XW, Zhang S, Chen MP, Hou HY, Wang ZL, Yu AL, Chen DH, Zhang XL, Hu JM, Dong YF, Liu D, Che RX (2023). The degradation of subalpine meadows significantly changed the soil microbiome. Agriculture, Ecosystems & Environment, 349, 108470. DOI: 10.1016/j.agee.2023.108470.
[18]	Luo G, Li W, Yan CL (2015). Research on the application of inactivated mycelium of Penicillium in rice production in Ning’er County. Yunnan Agricultural Science and Technology, 5, 14-16.
	[罗刚, 李伟, 闫春丽 (2015). 青霉菌灭活菌丝体在宁洱县水稻生产上的应用研究. 云南农业科技, 5, 14-16.]
[19]	McDonald JH (2014). Handbook of Biological Statistics. 3rd ed. Sparky House Publishing, Maryland, USA.
[20]	McSteen P (2009). Hormonal regulation of branching in grasses. Plant Physiology, 149, 46-55. DOI PMID
[21]	Rong CY, Liu YX, Chang ZY, Liu ZY, Ding YF, Ding CQ (2022). Cytokinin oxidase/dehydrogenase family genes exhibit functional divergence and overlap in rice growth and development, especially in control of tillering. Journal of Experimental Botany, 73, 3552-3568. DOI PMID
[22]	Ryu CM, Farag MA, Hu CH, Reddy MS, Wei HX, Paul W Paré PW, Kloepper JW (2003). Bacterial volatiles promote growth in Arabidopsis. Proceedings of the National Academy of Sciences of the United States of America, 100, 4927-4932.
[23]	Tomasi N, Weisskopf L, Renella G, Landi L, Pinton R, Varanini Z, Nannipieri P, Torrent J, Martinoia E, Cesco S (2008). Flavonoids of white lupin roots participate in phosphorus mobilization from soil. Soil Biology & Biochemistry, 40, 1971-1974.
[24]	Wang B, Liu H, Cai C, Thabit M, Wang P, Li G, Duan Z (2016). Effect of dry mycelium of Penicillium chrysogenum fertilizer on soil microbial community composition, enzyme activities and snap bean growth. Environmental Science and Pollution Research, 23, 20728-20738.
[25]	Wang B, Smith SM, Li J (2018). Genetic regulation of shoot architecture. Annual Review of Plant Biology, 69, 437-468. DOI PMID
[26]	Weisskopf L, Abou-Mansour E, Fromin N, Tomasi N, Santelia D, Edelkott I, Neumann G, Aragno M, Tabacchi R, Martinoia E (2006). White lupin has developed a complex strategy to limit microbial degradation of secreted citrate required for phosphate acquisition. Plant, Cell & Environment, 29, 919-927.
[27]	Wu XW (2019). Planting management and utilization techniques of Dabao Mountainous Grassland in the high cold mountain area of Zhaotong, Yunnan. Practical Rural Technology, (5), 45-47.
	[伍祥文 (2019). 云南昭通高寒山区大山包草地建植及管理利用技术. 农村实用技术, (5), 45-47.]
[28]	Xu CL, Xia KB, Zeng R, Yin SA, Wang XX, Zhang G, Ji GH, Chen SY (2009). Dry mycelium of Penicillium chrysogenum induces resistance against black shank and enhances growth of tobacco. Journal of Qinghai Normal University (Natural Science Edition), (2), 40-43.
	[徐长亮, 夏开宝, 曾嵘, 殷寿安, 王晓霞, 张鸽, 姬广海, 陈穗云 (2009). 青霉菌灭活菌丝体对烟草生长及黑胫病防治的影响. 青海师范大学学报(自然科学版), (2), 40-43.]
[29]	Yang CZ, Wang ZH, Guan XZ, Yang LX (2018). The effect of organic inducible disease inhibitor “Duotaibao” on the prevention and control of maize leaf spot disease. Yunnan Agriculture, (2), 77-78.
	[杨春震, 王忠华, 关学柱, 杨丽霞 (2018). 有机诱导抗病剂“多肽保”对玉米叶斑病的防控效果. 云南农业, (2), 77-78.]
[30]	Yang CZ, Wang ZH, Hu Y, He RF, Guan XZ, Yang LX (2017). The application of organic induced disease resistance agent “Duotaibao” in potato production. Yunnan Agriculture, (9), 55-56.
	[杨春震, 王忠华, 胡彦, 和汝凤, 关学柱, 杨丽霞 (2017). 有机诱导抗病剂“多肽保”在马铃薯生产上的应用. 云南农业, (9), 55-56.]
[31]	Zhang W, Peng KX, Cui FB, Wang DL, Zhao JZ, Zhang YJ, Yu NN, Wang YY, Zeng DL, Wang YH, Cheng ZK, Zhang KW (2021). Cytokinin oxidase/dehydrogenase OsCKX11 coordinates source and sink relationship in rice by simultaneous regulation of leaf senescence and grain number. Plant Biotechnology Journal, 19, 335-350. DOI PMID
[32]	Zhang XZ (1986). Determination of chlorophyll content in plants—Acetone ethanol mixture method. Liaoning Agricultural Science, (3), 26-28.
	[张宪政 (1986). 植物叶绿素含量测定——丙酮乙醇混合液法. 辽宁农业科学, (3), 26-28.]
[33]	Zhong Y, Chen ZZ, Xie H, Yan F, Shi HM, Zhang PY, Wang JG, Chen SY (2014). Primary researches on dry mycelium of Penicillium chrysogenum induced defense responses of tobacco BY-2 cell suspensions. Acta Phytopathologica Sinica, 44, 679-686.
	[钟宇, 陈壮壮, 谢虹, 燕飞, 时红敏, 张鹏远, 王建光, 陈穗云 (2014). 青霉菌灭活菌丝体诱导烟草BY-2悬浮细胞防卫反应的初步研究. 植物病理学报, 44, 679-686.] DOI
[34]	Zhong Y, Peng JJ, Chen ZZ, Xie H, Luo D, Dai JR, Yan F, Wang JG, Dong HZ, Chen SY (2015). Dry mycelium of Penicillium chrysogenum activates defense responses and restricts the spread of Tobacco Mosaic Virus in tobacco. Physiological and Molecular Plant Pathology, 92, 28-37.
[35]	Zhu H, Shen YX, Chen FJ, Fu X, Wang W (2022). Impacts of the degraded grassy hill and artificial reconstruction on soil seed bank in southern Zhaotong. Acta Agrestia Sinica, 30, 1359-1369. DOI
	[朱虹, 沈有信, 陈发军, 付珣, 王文 (2022). 昭通南方草山退化及人工改造对土壤种子库的影响. 草地学报, 30, 1359-1369.] DOI