Inoculation two azotobacter enhancing osmotic stress resistance and growth in wheat seedling

doi:10.3724/SP.J.1258.2013.00008

Abstract

Abstract:

Aims The seedling stage is the key stage of matter and energy accumulation in the wheat life cycle. Therefore, drought during the seedling stage affects population formation in late stages. In this study, wheat seedlings were inoculated with azotobacters Azorhizobium caulinodans ‘ORS571’ and Azospirillum brasilense ‘Yu62’.
Methods Wheat seedlings germination was screened in normal conditions and with PEG 6000 osmotic stress using seedlings inoculated with azotobacters. Root volume, relative water content (RWC), proline content and soluble protein content of seedling laminas were determined under PEG drought stress using seedlings inoculated with azotobacters on laminas.
Important findings The germination rate of wheat seedlings was significantly increased under drought stress when inoculated with azotobacters. Moreover, wheat seedlings inoculated with mixed azotobacters have more obvious growth promotion than when inoculated with a single azotobacter. The former laminas proline content, relative water content, proline content and soluble protein content had increased. The results showed that drought resistance of wheat seedlings was improved when inoculated with mixed azotobacters, which provided the foundation for further study of azotobacter-wheat interaction under drought stress.

Key words: drought resistance, mixed azotobacters, wheat seedling

LIU Hua-Wei, LIN Xiao-Jun, SUN Chao, LI Qiang, YANG Hu, GUO Ai-Guang. Inoculation two azotobacter enhancing osmotic stress resistance and growth in wheat seedling[J]. Chin J Plant Ecol, 2013, 37(1): 70-79.

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URL: https://www.plant-ecology.com/EN/10.3724/SP.J.1258.2013.00008

https://www.plant-ecology.com/EN/Y2013/V37/I1/70

Figures/Tables 12

Fig. 1 Growth curve of Azorhizobium caulinodans and Azospirillum brasilense under osmotic stress.

Table 1 Relative water content of wheat laminas under osmotic stress (mean ± SD)

小麦品种 Wheat cultivar	菌根处理 Mycorrhizal treatment	渗透胁迫 Osmotic stress (%)			复水 Rehydration (%)
小麦品种 Wheat cultivar	菌根处理 Mycorrhizal treatment	24 h	48 h	72 h	24 h	48 h
‘绵阳19’ ‘Mianyang 19’	不接种 No infected	97.44 ± 0.947^a	89.99 ± 2.326^b	86.99 ± 0.803^b	92.67 ± 1.042^b	96.56 ± 1.581^b
‘绵阳19’ ‘Mianyang 19’	接种 Infected	98.08 ± 1.782^a	96.87 ± 1.160^c	98.05 ± 0.949^e	98.02 ± 0.179^c	97.00 ± 0.399^b
‘西农979’ ‘Xinong 979’	不接种 No infected	95.68 ± 0.322^a	92.64 ± 2.056^bc	77.74 ± 1.738^a	94.46 ± 0.249^b	95.08 ± 1.499^ab
‘西农979’ ‘Xinong 979’	接种 Infected	95.67 ± 0.847^a	94.91 ± 2.992^c	90.79 ± 0.292^c	95.76 ± 0.020^bc	97.62 ± 1.876^b
‘周麦18’ ‘Zhoumai18’	不接种 No infected	97.28 ± 1.079^a	82.48 ± 1.067^a	74.67 ± 4.456^a	93.48 ± 0.800^b	96.27 ± 1.810^b
‘周麦18’ ‘Zhoumai18’	接种 Infected	97.28 ± 0.606^a	95.05 ± 1.959^c	90.22 ± 0.695^c	97.43 ± 2.094^c	98.07 ± 1.480^b
‘小偃22’ ‘Xiaoyan 22’	不接种 No infected	95.73 ± 0.371^a	84.01 ± 0.773^a	82.25 ± 1.723^b	87.78 ± 1.824^a	93.91 ± 0.292^a
‘小偃22’ ‘Xiaoyan 22’	接种 Infected	95.79 ± 0.713^a	97.86 ± 0.467^c	93.66 ± 0.834^d	97.05 ± 1.228^c	92.06 ± 1.093^a

Fig. 2 Germinate potential of wheat seeds infected by Azorhizobium caulinodans and Azospirillum brasilense under osmotic stress (mean ± SD). A, PBS positive control; B, negative control under osmotic stress; C, treatment of infection by Azorhizobium caulinodans under osmotic stress; D, treatment of infection by Azospirillum brasilense under osmotic stress.

Fig. 3 Germination percentage of wheat seeds infected by Azorhizobium caulinodans and Azospirillum brasilense under osmotic stress. A, PBS positive control; B, negative control under osmotic stress; C, treatment of infection by Azorhizobium caulinodans under osmotic stress; D, treatment of infection by Azospirillum brasilense under osmotic stress.

Fig. 4 Germination index of wheat seeds infected by Azorhizobium caulinodans and Azospirillum brasilense under osmotic stress. A, PBS positive control; B, negative control under osmotic stress; C, treatment of infection by Azorhizobium caulinodans under osmotic stress; D, treatment of infection by Azospirillum brasilense under osmotic stress.

Fig. 5 Effects of wheat seedling root length and plant height infected by Azorhizobium caulinodans and Azospirillum brasilense under osmotic stress (mean ± SD). A, infected by Azorhizobium caulinodans; B, infected by Azospirillum brasilense; C, infected by Azorhizobium caulinodans and Azospirillum brasilense; D, control.

Fig. 6 Root volume of wheat seedlings infected by Azorhizobium caulinodans and Azospirillum brasilense under osmotic stress (mean ± SD). A, infected by Azorhizobium caulinodans; B, infected by Azospirillum brasilense; C, infected by Azorhizobium caulinodans and Azospirillum brasilense; D, control.

Table 2 Colonization number of Azorhizobium caulinodans and Azospirillum brasilense in different wheat seedling organs after infected six days (three repeat)

	茎瘤固氮根瘤菌 Azorhizobium caulinodans			巴西固氮螺菌 Azospirillum brasilense
叶 Leaf (×10⁷)	14.2	11.9	12.1	16.4	15.2	14.2
根 Root (×10⁷)	2.4	1.9	2.1	1.8	2.0	1.8

Fig. 7 Colonization of green fluorescent protein (GFP)-Azorhizobium caulinodans in the wheat seeding via confocal laser scanning microscopy. A, Colonization of GFP-A. caulinodans in the wheat blade back, arrow point to stomas. B, Amplifying colonization of GFP-A. caulinodans in the middle of stoma. C, Colonization of GFP-A. caulinodans in the wheat stem vascular tissue. D, Colonization of GFP-A. caulinodans in the wheat root vascular tissue.

Fig. 8 Relative water content of wheat seedlings infected by Azorhizobium caulinodans and Azospirillum brasilense under osmotic stress (mean ± SD). A, infected by Azorhizobium caulinodans; B, infected by Azospirillum brasilense; C, infected by Azorhizobium caulinodans and Azospirillum brasilense; D, control.

Fig. 9 Proline content of wheat seedlings infected by Azorhizobium caulinodans and Azospirillum brasilense under osmotic stress (mean ± SD). A, infected by Azorhizobium caulinodans; B, infected by Azospirillum brasilense; C, infected by Azorhizobium caulinodans and Azospirillum brasilense; D, control.

Fig. 10 Soluble protein content of wheat seedlings infected by Azorhizobium caulinodans and Azospirillum brasilense under osmotic stress (mean ± SD). A, infected by Azorhizobium caulinodans; B, infected by Azospirillum brasilense; C, infected by Azorhizobium caulinodans and Azospirillum brasilense; D, control.

References 34

[1]	Athar M, Johnson DA (1996). Influence of drought on competition between selected Rhizobium meliloti strains and naturalized soil rhizobia in alfalfa. Plant and Soil, 184, 231-241.
[2]	Bushby HVA, Marshall KC (1977). Some factors affecting the survival of root-nodule bacteria on desiccation. Soil Biology & Biochemistry, 9, 143-147.
[3]	Cai H, Tang HX (2011). Agricultural drought assessment and on wheat growth impact. Modern Agricultural Sciences and Technology, (8), 292-293. (in Chinese)
	[ 蔡衡, 唐宏祥 (2011). 农业干旱的评估及对小麦生长的影响. 现代农业科技, (8), 292-293.]
[4]	Chen SY, Lang NJ, Li JY, Jia LQ, Wu LY, Mi FD (2004). Changes of leaf relative water content, relative plasma membrane permeability and proline content of seedlings of three species under drought stress. Journal of West China Forestry Science, 33(3), 30-33, 41. (in Chinese with English abstract)
	[ 陈少瑜, 郎南军, 李吉跃, 贾利强, 吴丽圆, 米方佃 (2004). 干旱胁迫下3树种苗木叶片相对含水量、质膜相对透性和脯氨酸含量的变化. 西部林业科学, 33(3), 30-33, 41.]
[5]	Chi F (2006). Migration of Rhizobia in Plants and Proteome Analysis of Their Interaction. PhD dissertation, Institute of Botany, Chinese Academy of Sciences, Beijing. (in Chinese with English abstract)
	[ 迟峰 (2006). 根瘤菌在植物内的迁移运动及其与植物相互作用的蛋白质组学研究. 博士学位论文, 中国科学院植物研究所, 北京.]
[6]	Chi F, Shen SH, Cheng HP, Jing YX, Yanni YG, Dazzo FB (2005). Ascending migration of endophytic rhizobia, from roots to leaves, inside rice plants and assessment of benefits to rice growth physiology. Applied and Environmental Microbiology, 71, 7271-7278. URL PMID
[7]	de Oliveira AC, Varshney RK (2011). Root Genomics. Springer, New York. 1-10.
[8]	Döbereiner J, Day JM, Dart PJ (1972). Nitrogenase activity in the rhizosphere of sugar cane and some other tropical grasses. Plant and Soil, 37, 191-196.
[9]	Fu B, Wang WW, Tang M, Chen XD (2009). Isolation and identification of hydrogen-oxidizing bacteria producing 1-aminocyclopropane-1- carboxylate deaminase and the determination of enzymatic activity. Acta Microbiologica Sinica, 49, 395-399. (in Chinese with English abstract) URL PMID
	[ 付博, 王卫卫, 唐明, 陈兴都 (2009). 一株产1-氨基环丙烷-1-羧酸脱氨酶的氢氧化细菌的分离鉴定及酶活力测定及酶活力测定. 微生物学报, 49, 395-399.] PMID
[10]	Fuhrmann J, Davey CB, Wollum AG (1986). Desiccation tolerance of clover rhizobia in sterile soils. Soil Science Society of America Journal, 50, 639-644.
[11]	Glick BR, Bashan Y (1997). Genetic manipulation of plant growth-promoting bacteria to enhance biocontrol of phytopathogens. Biotechnology Advances, 15, 353-378. URL PMID
[12]	Guo AG, Guo ZK (2007). Biochemical Experimental Technology. Higher Education Press, Beijing. (in Chinese)
	[ 郭蔼光, 郭泽坤 (2007). 生物化学实验技术. 高等教育出版社, 北京.]
[13]	Ji KX, Chi F, Yang MF, Shen SH, Jing YX, Dazzo FB, Cheng HP (2010). Movement of rhizobia inside tobacco and lifestyle alternation from endophytes to free-living rhizobia on leaves. Journal of Microbiology and Biotechnology, 20, 238-244. URL PMID
[14]	Kang YJ, Cheng J, Mei LJ, Hu J, Piao Z, Yin SX (2010). Action mechanisms of plant growth-promoting rhizobacteria (PGPR): a review. Chinese Journal of Applied Ecology, 21, 232-238. (in Chinese with English abstract) URL PMID
	[ 康贻军, 程洁, 梅丽娟, 胡健, 朴哲, 殷士学 (2010). 植物根际促生菌作用机制研究进展. 应用生态学报, 21, 232-238.] PMID
[15]	Kase H, Kochler M, Stuzel H (2004). Root growth and dry matter partitioning of cauliﬂower under drought stress conditions: measurement and simulation. European Journal of Agronomy, 20, 379-394.
[16]	Kozlowski TT, Pallardy SG (2002). Acclimation and adaptive responses of woody plants to environmental stresses. The Botanical Review, 68, 270-334.
[17]	Li XW, Li JM, Duan LS, Li ZH (2010). Primary study on inducing effect of coronatine on drought tolerance of winter wheat seedlings. Journal of Triticeae Crops, 30, 676-679. (in Chinese with English abstract)
	[ 李相文, 李建民, 段留生, 李召虎 (2010). 冠菌素诱导冬小麦幼苗抗旱性的初步研究. 麦类作物学报, 30, 676-679.]
[18]	Li YG, Zhou JC (2002). Root colonization of Sinorhizobium fredii pSym-cured strain HN01 in rhizosphere of Glycine max. Acta Ecologica Sinica, 22, 1420-1424. (in Chinese with English abstract)
	[ 李友国, 周俊初 (2002). 消除共生质粒的费氏中华根瘤菌HND29SR在大豆根圈的定殖动态. 生态学报, 22, 1420-1424.]
[19]	Liang Y, Chen SP, Gao YB, Ren AZ. (2002). Effects of endophyte infection on the growth of Lolium perenne L. under drought stress. Acta Phytoecologica Sinica, 26, 621-626. (in Chinese with English abstract)
	[ 梁宇, 陈世苹, 高玉葆, 任安芝 (2002). 内生真菌感染对干旱胁迫下黑麦草生长的影响. 植物生态学报, 26, 621-626.]
[20]	Liu HF, Gao YB, Zhang Q, Li X, Li CL (2004). Physio- ecological responses and their adaptation of different geographic Leymus chinensis populations to soil drought stress. Acta Scientiarum Naturalium Universitatis Nankaiensis (Natural Science Edition), 37(4), 105-110. (in Chinese with English abstract)
	[ 刘慧芳, 高玉葆, 张强, 李欣, 李长林 (2004). 不同种群羊草幼苗对土壤干旱胁迫的生理生态响应. 南开大学学报(自然科学版), 37(4), 105-110.]
[21]	Liu HW, Sun C, Yang H, Lin XJ, Guo AG (2012). Promotion for wheat growth and root colonization after infecting wheat seeds with Azorhizobium caulinodans. Plant Nutrition and Fertilizer Science, 18, 210-217. (in Chinese with English abstract) DOI URL
	[ 刘华伟, 孙超, 杨呼, 林晓军, 郭蔼光 (2012). 田菁茎瘤固氮根瘤菌对小麦种子侵染的促生作用及其在根系内的定殖. 植物营养与肥料学报, 18, 210-217.]
[22]	Liu HW, Wang QH, Zhang H, Wang R, Xiao HL, Guo AG (2009). Colonization of Azospirillum brasilense Yu62 in wheat via EGFP. Acta Botanica Boreali-Occidentalia Sinica, 29, 2367-2372. (in Chinese with English abstract)
	[ 刘华伟, 王庆贺, 张宏, 王蕊, 肖红利, 郭蔼光 (2009). 巴西固氮螺菌Yu62的EGFP标记及其在小麦体内的定殖研究. 西北植物学报, 29, 2367-2372.]
[23]	Lu YH, Zhang FS (2006). The advances in rhizosphere microbiology. Soils, 38, 113-121. (in Chinese with English abstract)
	[ 陆雅海, 张福锁 (2006). 根际微生物研究进展. 土壤, 38, 113-121.]
[24]	Pan PP, Zhou HB (1995). Plant hormones produced by Azorhizobium caulinodans ORS 571. Microbiology, 22, 10-13. (in Chinese with English abstract)
	[ 潘佩平, 周鸿宾 (1995). 茎瘤固氮根瘤菌(Azorhizobium caulinodans) ORS571产生的植物激素. 微生物学通报, 22, 10-13.]
[25]	Shen SH, Jing YX (2003). Present and prospect of nitrogen-fixing in China. Chinese Science Bulletin, 48, 535-540. (in Chinese)
	[ 沈世华, 荆玉祥 (2003). 中国生物固氮研究现状和展望. 科学通报, 48, 535-540.]
[26]	Shi Y, Lin Q, Wei DB, Yu ZW (1996). Impacts of soil water stress on photosynthesis and yield of winter wheat. Acta Agriculturae Boreali- Sinica, 11(4), 80-83. (in Chinese with English abstract)
	[ 石研, 林琪, 魏东斌, 于振文 (1996). 土壤水分胁迫对冬小麦光合及产量的影响. 华北农学报, 11(4), 80-83.]
[27]	Timmusk S, Wagner EGH (1999). The plant growth-promoting rhizobacterium Paenibacillus polymyxa induces changes in Arabidopsis thaliana gene expression: a possible connection between biotic and abiotic stress responses. Molecular Plant-Microbe Interactions, 12, 951-959. URL PMID
[28]	Wang WE, Li Y, Su N, Yu WC (2011). The effects of salt stress on seed germination of Indigofera amblyantha. Hubei Agricultural Sciences, 50, 321-324. (in Chinese with English abstract)
	[ 王文恩, 李颖, 苏农, 余文成 (2011). 盐胁迫对多花木蓝种子萌发的影响. 湖北农业科学, 50, 321-324.]
[29]	Xu XL, Guan XQ, Liu GS (2003). Effects of cooperation between associative azotobacteria and rhizobia on wheat seedling. Chinese Journal of Eco-Agriculture, 11(3), 66-68. (in Chinese with English abstract)
	[ 徐兴良, 关秀清, 刘公社 (2003). 联合固氮菌与根瘤菌协同作用对小麦幼苗的影响. 中国生态农业学报, 11(3), 66-68.]
[30]	Ye AH, Yuan Y, Yang L, Cai YP, Tian SN (2003). A comparison of the effects of two arbuscular mycorrhizal fungal species on photosynthesis, transpiration and water use efficiency of wheat. Chinese Agricultural Science Bulletin, 19(3), 18-20. (in English with Chinese abstract)
	[ 叶爱华, 袁艺, 杨莉, 蔡永萍, 田胜尼 (2003). 两种丛枝菌根菌抗旱效应的比较. 中国农学通报, 19(3), 18-20.]
[31]	Yu CQ (2011). China’s water crisis needs more than words. Nature, 470, 307. DOI URL PMID
[32]	Zhang DZ, Wang PH, Zhao HX (1990). Determination of the content of free proline in wheat leaves. Plant Physiology Communications, (4), 62-65. (in Chinese with English abstract)
	[ 张殿忠, 王沛洪, 赵会贤 (1990). 测定小麦叶片游离脯氨酸含量的方法. 植物生理学通讯, (4), 62-65.]
[33]	Zhao SJ, Li SL, Wang ZQ (2004). Effects of VA mycorrhizal fungi on drought resistance of spring wheat under non-irrigation. Journal of Inner Mongolia Institute of Agriculture and Animal Husbandry, 25(4), 43-46. (in Chinese with English abstract)
	[ 赵士杰, 李树林, 王志强 (2004). VA菌根真菌对旱作春小麦抗旱性的影响. 内蒙古农业大学学报, 25(4), 43-46.]
[34]	Zhou LP, Liu WJ, Ma HC, Wu JR (2010). Research progress on drought resistance of rhizobium strains and plants inoculated with rhizobium. Journal of Anhui Agricultural Sciences, 38, 11978-11980, 11983. (in Chinese with English abstract)
	[ 周利平, 刘文杰, 马焕成, 伍建榕 (2010). 根瘤菌菌株和接种根瘤菌植株的耐旱性研究进展. 安徽农业科学, 38, 11978-11980, 11983.]