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作物学报 ›› 2025, Vol. 51 ›› Issue (2): 358-369.doi: 10.3724/SP.J.1006.2025.44091

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

嗜线虫沙雷氏菌Serratia nematodiphila TG10增强油菜耐盐碱能力

苏晴芳1,2(), 孙小钊1,2, 林杨1, 付艳苹1, 程家森1, 谢甲涛1,2, 姜道宏1,2, 陈桃1,2,*()   

  1. 1华中农业大学植物科学技术学院 / 农业微生物资源发掘与利用全国重点实验室, 湖北武汉 430070
    2湖北洪山实验室, 湖北武汉 430070
  • 收稿日期:2024-06-07 接受日期:2024-09-18 出版日期:2025-02-12 网络出版日期:2024-10-10
  • 通讯作者: 陈桃, E-mail: taochen@mail.hzau.edu.cn
  • 作者简介:E-mail: suqingfang@webmail.hzau.edu.cn
  • 基金资助:
    中央高校基本科研专项资金项目(X2662024ZKPY004);中央高校基本科研专项资金项目(2662023PY006);财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-12)

Serratia nematodiphila TG10 enhanced salt-alkali tolerance in rapeseed

SU Qing-Fang1,2(), SUN Xiao-Zhao1,2, LIN Yang1, FU Yan-Ping1, CHENG Jia-Sen1, XIE Jia-Tao1,2, JIANG Dao-Hong1,2, CHEN Tao1,2,*()   

  1. 1College of Plant Science and Technology, Huazhong Agricultural University / National Key Laboratory of Agricultural Microbiology, Wuhan 430070, Hubei, China
    2Hubei Hongshan Laboratory, Wuhan 430070, Hubei, China
  • Received:2024-06-07 Accepted:2024-09-18 Published:2025-02-12 Published online:2024-10-10
  • Contact: E-mail: taochen@mail.hzau.edu.cn
  • Supported by:
    Research Funds for the Central Universities(X2662024ZKPY004);Research Funds for the Central Universities(2662023PY006);China Agriculture Research System of MOF and MARA(CARS-12)

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摘要：

油菜具有较强的耐盐碱能力, 本研究从天津盐碱土中分离得到一株嗜线虫沙雷氏菌Serratia nematodiphila TG10, 它不仅能够在8% NaCl和pH 10.15的R2A培养基上生长, 还能定殖在油菜根系和根内。在盐胁迫条件下, TG10菌株处理后可以促进拟南芥和油菜生长。含盐1.2%基质土和哈尔滨盐碱土盆栽试验中, TG10处理后可以促进油菜鲜重和干重的增加, Na+含量显著下降, K+与Na+的含量比值增加; 同样在天津盐碱土和吉林盐碱土盆栽试验中, TG10处理后均可显著增加油菜鲜重、叶绿素和脯氨酸的含量。吉林盐碱土盆栽油菜的转录组数据表明, TG10菌株处理后细胞色素P450代谢通路、硫代葡萄糖硫苷生物通路等显著富集, 一些抗逆相关的基因显著上调表达。另外, TG10菌株能抑制油菜病原菌的生长, 诱导油菜抗菌核病和灰霉病。本研究结果表明, 嗜线虫沙雷氏菌S. nematodiphila TG10能增强油菜在不同类型盐碱土中的耐盐能力, 为盐碱地的生物修复提供微生物菌种资源和理论支撑。

关键词: 盐碱土, 油菜, 嗜线虫沙雷氏菌, 耐盐碱机制, 诱导抗性

Abstract:

Rapeseed exhibits strong resistance to salt-alkali stress. In this study, a strain of Serratia nematodiphila TG10 was isolated from saline-alkali soil in Tianjin. This strain can not only grow on a 6% NaCl R2A medium plate with a pH of 10.15 but can also colonize the roots and rhizosphere of rapeseed. Under salt stress, the TG10 strain promoted the growth of both Arabidopsis and rapeseed. In pot experiments using a 1.2% salt-containing matrix soil and saline-alkali soil from Harbin, TG10 treatment enhanced the fresh and dry weights of rapeseed, reduced Na? content, and increased the K?/Na? ratio. In experiments with saline-alkali soils from Tianjin and Jilin, TG10 treatment significantly increased the fresh weight, chlorophyll content, and proline levels in rapeseed. Transcriptome analysis of rapeseed grown in Jilin saline-alkali soil revealed that the cytochrome P450 metabolic pathway and glucosinolate biosynthesis pathway were significantly enriched following TG10 treatment, with significant upregulation of several stress-related genes. Additionally, the TG10 strain inhibited the growth of pathogenic bacteria in rapeseed and induced resistance against Sclerotinia sclerotiorum and Botrytis cinerea. These findings suggest that S. nematodiphila TG10 can enhance the salt tolerance of rapeseed in various types of saline-alkali soils, providing a valuable resource and theoretical foundation for the biorefining of microorganisms in saline-alkali environments.

Key words: saline-alkali soil, rapeseed, Serratia nematodiphila, saline-alkali resistance mechanism, induced resistance

表1

3种不同类型盐碱土理化性质"

盐碱土来源
Saline-alkali soil source		 有机质
Organic matter
(g kg-1)		 电导率
Electrical
conductivity
(mS cm-1)		 有效磷
Available
phosphorus
(mg kg-1)		 速效钾
Available
potassium
(mg kg-1)		 钠离子
Sodium ion
(mg kg-1)		 pH
天津 Tianjin 13.73±0.42 13.58±0.27 53.73±1.04 332.23±22.33 1389.09±45.72 8.97±0.08
吉林 Jilin 19.13±5.58 6.92±0.43 7.88±1.26 230.02±31.83 2294.10±17.92 10.05±0.04
黑龙江哈尔滨
Harbin, Heilongjiang		 9.35±1.10 0.61±0.19 16.37±1.15 229.00±6.08 941.82±243.85 10.09±0.07

图1

嗜线虫沙雷氏菌TG10的鉴定及油菜根系定殖能力测试 A: 菌株TG10在不同NaCl含量以及pH的R2A培养基生长3 d的菌落形态; B: 根据菌株TG10的16S全长, 使用MEGA 7.0软件中的Neighbor-Joining构建系统发育树; C, D: TG10处理油菜, 无菌培养14 d后, 根系和根内分离的TG10的表型(C)和菌落数目统计(D)。每一株油菜的所有根系为一个样本, 3个生物学重复。图中误差线为平均值±标准差, 数据均使用t-test进行检验分析, *表示P < 0.05。"

图2

菌株TG10增强拟南芥和油菜在无菌/半无菌条件下的耐盐能力 A~C: 在100 mmol L-1 NaCl胁迫下, 拟南芥种子与菌株共培养14 d后的表型(A)、鲜重(B)和根长(C)统计, n = 37~64; D~F: TG10增强油菜在水培条件下的耐盐能力, 菌液处理的油菜幼苗在含75 mmol L-1 NaCl的霍格兰营养液中水培7 d的表型(D)、鲜重(E)和根长(F)的统计, n = 15~26。图中误差线为平均值±标准差, 标尺为1 cm。数据均采用t检验分析, *表示P < 0.05, **表示P < 0.01, ***表示P < 0.001, ****表示P < 0.0001。"

图3

菌株TG10处理减低油菜的Na+含量 A~F: 在1.2% NaCl基质土壤中, 油菜接种菌液28 d后的表型(A)、鲜重(B)、干重(C)、地上部分Na+含量(D)、地上部分K+含量(E)和K+/Na+含量的比值(F), A中标尺为2 cm, B中n = 12~14, C中n = 7, D、E和F中n = 3。G~L: 在哈尔滨盐碱土中, 用细菌处理28 d后的油菜表型(G)、鲜重(H)、干重(I)和地上部分Na+含量(J)、地上部分K+含量(K)和K+/Na+含量的比值(L), G中标尺为2 cm, H中n = 13~15, I中n = 9~10, J、K和L中n = 3。图中误差线为平均值±标准差, 数据均采用t检验分析, *表示P < 0.05, **表示P < 0.01, ***表示P < 0.001, ****表示P < 0.0001。"

图4

菌株TG10显著提高油菜在盐碱土中叶绿素和脯氨酸的含量 A~D: 天津盐碱土盆栽试验, 菌液处理28 d后的油菜植株表型(A)、地上部分鲜重(B)、叶绿素含量(C)和脯氨酸含量(D)的测定, A中标尺为4 cm, B中n = 15, C中n = 3, D中n = 3~5。E~H: 吉林盐碱土盆栽试验, 菌液处理28 d后的油菜表型(E)、鲜重(F)、叶绿素含量(G)和脯氨酸含量(H)的测定, E中标尺为2 cm, F中n = 28, G中n = 5, H中n = 3~4。图中误差线为平均值±标准差, 数据均采用t检验分析, *表示P < 0.05, **表示P < 0.01, ***表示P < 0.001, ****表示P< 0.0001。"

图5

盐胁迫下菌株TG10诱导基因差异表达 A: 将处理组TG10与对照组CK进行基因相对表达量的比较, 随后将获得的DEGs进行火山图的绘制, 蓝色表示下调基因(log2 (FC) < -1, P < 0.05), 橘色表示上调基因(log2 (FC) > 1, P < 0.05), 灰色表示非显著变化基因。B: 将TG10 vs CK的上调DEGs进行GO富集, 选取不重复的丰度前20的条目绘制气泡图, 图中主要展示基因参与的生物学过程(Biological process, BP)。C: 将TG10 vs CK的上调DEGs进行KEGG富集, 选取P < 0.05的通路绘制气泡图。D: 选取KEGG中与抗逆相关通路的35个基因进行热图的绘制。"

图6

菌株TG10诱导油菜抵御核盘菌和灰霉菌的侵染 菌株TG10划线于核盘菌(A)和灰霉菌(B)的四周, 两者在PDA平板上生长2~4 d的表型及统计的真菌直径, 标尺为1 cm, n = 3; 菌液处理的油菜子叶上接种核盘菌(C)和灰霉菌(D)的菌丝块36 h后的表型及统计的病斑大小, 标尺为0.5 cm, n = 6。图中误差线为平均值±标准差, 数据均采用t-test检验分析, *表示P < 0.05, **表示P < 0.01。"

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