接种两种固氮菌增强小麦幼苗抗渗透胁迫及生长能力

doi:10.3724/SP.J.1258.2013.00008

摘要/Abstract

摘要：

苗期是小麦(Triticum aestivum)物质和能量积累的关键时期, 苗期干旱影响小麦的后期群体建成。利用田菁茎瘤固氮根瘤菌(Azorhizobium caulinodans) ‘ORS571’与巴西固氮螺菌(Azospirillum brasilense) ‘Yu62’浸种侵染小麦和聚乙二醇(PEG)模拟渗透胁迫, 研究渗透胁迫下接菌小麦种子的发芽状况; 利用固氮菌涂抹小麦幼苗叶部, 测定PEG模拟渗透胁迫下小麦幼苗根体积、叶片相对含水量、脯氨酸含量及可溶性蛋白含量, 探究固氮菌增强小麦幼苗抗渗透胁迫的能力。结果表明, 接种混合固氮菌后在渗透胁迫下小麦种子的发芽率明显提高; 在渗透胁迫下叶部涂抹固氮菌小麦的根体积、叶片相对含水量、脯氨酸含量及可溶性蛋白含量明显升高, 表明接种固氮菌可提高小麦幼苗的抗渗透胁迫能力, 且混合固氮菌对小麦幼苗叶片的增强效果优于单一固氮菌。

关键词: 抗旱性, 混合固氮菌, 小麦幼苗

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

刘华伟, 林晓军, 孙超, 李强, 杨呼, 郭蔼光. 接种两种固氮菌增强小麦幼苗抗渗透胁迫及生长能力. 植物生态学报, 2013, 37(1): 70-79. DOI: 10.3724/SP.J.1258.2013.00008

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. Chinese Journal of Plant Ecology, 2013, 37(1): 70-79. DOI: 10.3724/SP.J.1258.2013.00008

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链接本文: https://www.plant-ecology.com/CN/10.3724/SP.J.1258.2013.00008

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

图/表 12

图1 渗透胁迫下田菁茎瘤固氮根瘤菌和巴西固氮螺菌的生长曲线。

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

表1 渗透胁迫下小麦叶片的相对含水量(平均值±标准偏差)

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

图2 渗透胁迫下小麦种子接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌的发芽势(平均值±标准偏差)。A, PBS正对照; B, 渗透胁迫处理负对照; C, 渗透胁迫下接种田菁茎瘤固氮根瘤菌; D, 渗透胁迫下接种巴西固氮螺菌。

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.

图3 渗透胁迫小麦种子接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌的发芽率。A, PBS正对照; B, 渗透胁迫处理负对照; C, 渗透胁迫下接种田菁茎瘤固氮根瘤菌; D, 渗透胁迫下接种巴西固氮螺菌。

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.

图4 渗透胁迫下小麦种子接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌的发芽指数。A, PBS正对照; B, 渗透胁迫处理负对照; C, 渗透胁迫下接种田菁茎瘤固氮根瘤菌; D, 渗透胁迫下接种巴西固氮螺菌。

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.

图5 渗透胁迫下接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌对小麦幼苗根长与株高的影响(平均值±标准偏差)。A, 接种田菁茎瘤固氮根瘤菌; B,接种巴西固氮螺菌; C, 接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌; D, 对照。

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.

图6 渗透胁迫下叶片接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌小麦幼苗的根体积(平均值±标准偏差)。A, 接种田菁茎瘤固氮根瘤菌; B,接种巴西固氮螺菌; C, 接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌; D, 对照。

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.

表2 接种6天后不同小麦幼苗器官中田菁茎瘤固氮根瘤菌和巴西固氮螺菌定殖数目(3次重复)

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

图7 激光共聚焦显微镜检测绿色荧光蛋白(GFP)-田菁茎瘤固氮根瘤菌在小麦体内的定殖。A, GFP-田菁茎瘤固氮根瘤菌吸附在小麦的叶背面, 箭头所示为分布气孔处的GFP-田菁茎瘤固氮根瘤菌。B, GFP-田菁茎瘤固氮根瘤菌定殖在气孔处的局部放大照片。C, GFP-田菁茎瘤固氮根瘤菌定殖在小麦茎的维管组织中。D, GFP-田菁茎瘤固氮根瘤菌定殖在小麦根的维管组织中。

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.

图8 渗透胁迫下叶片接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌小麦幼苗的相对含水量(平均值±标准偏差)。A, 接种田菁茎瘤固氮根瘤菌; B,接种巴西固氮螺菌; C, 接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌; D, 对照。

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.

图9 渗透胁迫下叶片接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌小麦幼苗的脯氨酸含量(平均值±标准偏差)。A, 接种田菁茎瘤固氮根瘤菌; B,接种巴西固氮螺菌; C, 接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌; D, 对照。

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.

图10 渗透胁迫下叶片接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌小麦幼苗的可溶性蛋白含量(平均值±标准偏差)。A, 接种田菁茎瘤固氮根瘤菌; B,接种巴西固氮螺菌; C, 接种田菁茎瘤固氮根瘤菌和巴西固氮螺菌; D, 对照。

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.

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