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作物学报 ›› 2024, Vol. 50 ›› Issue (8): 2122-2130.doi: 10.3724/SP.J.1006.2024.44006

• 研究简报 • 上一篇    下一篇

大豆萌发期耐盐性鉴定方法建立及耐盐大豆资源筛选

刘欣玥1,2(), 郭潇阳2, 王欣茹2, 辛大伟1,*(), 关荣霞2,*(), 邱丽娟2,*()   

  1. 1东北农业大学农学院, 黑龙江哈尔滨 150030
    2农作物基因资源与遗传改良国家重大科学工程 / 农业农村部北京大豆生物学重点实验室/作物基因资源与育种全国重点实验室 / 中国农业科学院作物科学研究所, 北京 100081
  • 收稿日期:2024-01-09 接受日期:2024-04-01 出版日期:2024-08-12 网络出版日期:2024-04-20
  • 通讯作者: * 辛大伟, E-mail: xdawei@163.com;关荣霞, E-mail: guanrongxia@caas.cn;邱丽娟, E-mail: qiulijuan@caas.cn
  • 作者简介:E-mail: 1240054143@qq.com
  • 基金资助:
    科技创新2030-重大项目(2023ZD0403601);国家自然科学基金项目(31830066)

Establishment of screening method for salt tolerance at germination stage and identification of salt-tolerant germplasms in soybean

LIU Xin-Yue1,2(), GUO Xiao-Yang2, WANG Xin-Ru2, XIN Da-Wei1,*(), GUAN Rong-Xia2,*(), QIU Li-Juan2,*()   

  1. 1Agricultural College, Northeast Agricultural University, Harbin 150030, Heilongjiang, China
    2National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI) / Key Laboratory of Soybean Biology, Ministry of Agriculture and Rural Affairs / State Key Laboratory of Crop Gene Resources and Breeding / Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2024-01-09 Accepted:2024-04-01 Published:2024-08-12 Published online:2024-04-20
  • Contact: * E-mail: xdawei@163.com;E-mail: guanrongxia@caas.cn;E-mail: qiulijuan@caas.cn
  • Supported by:
    Science and Technology Innovation 2030-Major Projects(2023ZD0403601);National Natural Science Foundation of China(31830066)

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摘要：

土壤盐渍化会影响大豆萌发和生长, 筛选萌发期耐盐大豆种质对于耐盐大豆品种选育及盐渍土利用意义重大。本研究以8份代表性大豆种质为材料, 蛭石为培养基质, 用150 mmol L-1 NaCl进行盐胁迫处理, 以水处理作为对照, 测量2 d和3 d的大豆发芽率和芽长, 旨在建立大豆萌发期耐盐性鉴定指标和评价方法。与对照相比, 盐胁迫下大豆发芽率和芽长均下降。盐处理2 d不同种质间相对发芽率、相对芽长无显著差异, 盐处理3 d相对芽长差异显著, 可用以进行不同大豆萌发期耐盐性评价。以150 mmol L-1 NaCl对48份大豆种质进行盐处理, 3 d相对芽长作为评价指标, 将萌发期耐盐性划分为5级, 1级为高度耐盐型, 2级为耐盐型, 3级为中度盐敏感型, 4级为盐敏感型, 5级为高度盐敏感型。鉴定出萌发期高度耐盐种质5份, 耐盐种质12份。同时对48份种质进行出苗期和苗期耐盐性鉴定, 鉴定出3份从萌发期到苗期均耐盐的种质, 分别为中黄685、黑农94和东生104。本研究建立了一套室内大豆萌发期耐盐性鉴定方法, 为耐盐种质鉴定利用奠定了基础。

关键词: 大豆, 种质资源, 萌发期, 耐盐性鉴定

Abstract:

Salinity tolerance is prioritized for soybean because salt can affect soybean germination and growth. Screening for salt-tolerant soybean germplasms is of great significance for the breeding salt-tolerant soybean varieties and the utilization of saline lands for production. In this study, eight representative soybean germplasms were used as the experimental materials, soybean seeds were planted in vermiculite and treated with 150 mmol L-1 NaCl or water as the control. The germination rate and radicle length of soybean were measured after 2 days and 3 days treatment, aiming to establish the identification index and evaluation method of soybean salt tolerance at germination stage. In comparison to control, the germination rate and radicle length of soybean decreased under salt stress. No significant difference was observed in the germination rate and relative radicle length (RRL) between germplasms after 2 days salt treatment. Significant difference was observed in RRL after 3 days salt stress. Therefore, RRL was selected as the evaluation indicator to distinguish salt tolerance of 48 soybean germplasms. The 48 germplasms were divided into 5 levels by using RRL after 3 days salt treatment. Level 1, level 2, level 3, level 4, and level 5 was high tolerant, tolerant, medium sensitive, sensitive, and most sensitive, respectively. Five high salt-tolerant germplasms and 12 salt-tolerant germplasms were identified at germination stage. These germplasms were screened for salt tolerance at both emergence and seedling stages Three soybeans were identified for salt tolerance from germination stage to seeding stage, namely Zhonghuang 685, Heinong 94, and Dongsheng 104. In conclusion, a screening method for salt tolerance at germination stage under controlled condition was established, which laid a foundation for the identification and utilization of salt tolerant germplasms.

Key words: soybean, germplasm, germination stage, evaluation for salt tolerance

表1

48个大豆种质萌发期、出苗期和苗期耐盐指标及GmSALT3单倍型"

序号
No.		 大豆名称
Soybean name		 3 d相对芽长Relative radicle length 3 d after salt stress 耐盐等级Salt tolerance level GmSALT3基因型
Haplotype of GmSALT3
萌发期Germination stage 出苗期Emergence stage 苗期Seedling stage
1 中黄685 Zhonghuang 685 0.61 1 1 2 耐盐单倍型 Salt-tolerant haplotype
2 东生104 Dongsheng 104 0.60 1 2 2 耐盐单倍型 Salt-tolerant haplotype
3 黑农94 Heinong 94 0.59 1 2 1 耐盐单倍型 Salt-tolerant haplotype
4 通农15 Tongnong 15 0.58 1 3 2 耐盐单倍型 Salt-tolerant haplotype
5 铁丰31 Tiefeng 31 0.56 1 2 4 敏感单倍型 Salt-sensitive haplotype
6 中吉602 Zhongji 602 0.54 2 1 3 敏感单倍型 Salt-sensitive haplotype
7 吉科黄豆20 Jikehuangdou 20 0.53 2 5 2 耐盐单倍型 Salt-tolerant haplotype
8 赤16-33 Chi 16-33 0.50 2 4 3 敏感单倍型 Salt-sensitive haplotype
9 中黄39 Zhonghuang 39 0.50 2 2 3 敏感单倍型 Salt-sensitive haplotype
10 齐农7号 Qinong 7 0.50 2 4 4 敏感单倍型 Salt-sensitive haplotype
11 通农943 Tongnong 943 0.50 2 2 5 敏感单倍型 Salt-sensitive haplotype
12 中黄320 Zhonghuang 320 0.49 2 2 4 敏感单倍型 Salt-sensitive haplotype
13 中黄324 Zhonghuang 324 0.48 2 1 4 耐盐单倍型 Salt-tolerant haplotype
14 中联豆5022 Zhongliandou 5022 0.48 2 4 4 敏感单倍型 Salt-sensitive haplotype
15 汾豆78 Fendou 78 0.47 2 3 1 耐盐单倍型 Salt-tolerant haplotype
16 中黄204 Zhonghuang 204 0.47 2 3 1 耐盐单倍型 Salt-tolerant haplotype
17 长农52 Changnong 52 0.46 2 3 4 敏感单倍型 Salt-sensitive haplotype
18 抗线虫8号 Kangxianchong 8 0.45 3 2 4 敏感单倍型 Salt-sensitive haplotype
19 中黄343 Zhonghuang 343 0.44 3 1 5 敏感单倍型 Salt-sensitive haplotype
20 中黄48 Zhonghuang 48 0.44 3 1 4 敏感单倍型 Salt-sensitive haplotype
21 中黄349 Zhonghuang 349 0.44 3 3 1 耐盐单倍型 Salt-tolerant haplotype
22 Williams 82 0.43 3 3 3 敏感单倍型 Salt-sensitive haplotype
23 中黄311 Zhonghuang 311 0.41 3 1 4 敏感单倍型 Salt-sensitive haplotype
24 通农102 Tongnong 102 0.40 3 2 4 敏感单倍型 Salt-sensitive haplotype
25 中黄314 Zhonghuang 314 0.40 3 1 1 耐盐单倍型 Salt-tolerant haplotype
26 中黄329 Zhonghuang 329 0.40 3 2 4 敏感单倍型 Salt-sensitive haplotype
27 中黄354 Zhonghuang 354 0.40 3 1 1 耐盐单倍型 Salt-tolerant haplotype
28 公552 Gong 552 0.39 3 4 4 敏感单倍型 Salt-sensitive haplotype
29 中黄357 Zhonghuang 357 0.39 3 1 2 耐盐单倍型 Salt-tolerant haplotype
30 吉大豆2号 Jidadou 2 0.37 3 5 2 耐盐单倍型 Salt-tolerant haplotype
31 中黄37 Zhonghuang 37 0.36 3 5 1 耐盐单倍型 Salt-tolerant haplotype
32 中黄306 Zhonghuang 306 0.36 3 3 1 耐盐单倍型 Salt-tolerant haplotype
33 中黄339 Zhonghuang 339 0.36 3 1 1 耐盐单倍型 Salt-tolerant haplotype
34 吉育3528 Jiyu 3528 0.35 4 1 3 敏感单倍型 Salt-sensitive haplotype
35 中黄301 Zhonghuang 301 0.34 4 2 3 敏感单倍型 Salt-sensitive haplotype
36 通野19-4260-3 Tongye 19-4260-3 0.33 4 5 4 敏感单倍型 Salt-sensitive haplotype
37 吉农169 Jinong 169 0.32 4 4 3 杂合 Heterozygous haplotype
38 齐农5号 Qinong 5 0.32 4 4 3 敏感单倍型 Salt-sensitive haplotype
39 黑农532 Heinong 532 0.32 4 4 4 敏感单倍型 Salt-sensitive haplotype
40 NY27-38 0.31 4 4 2 耐盐单倍型 Salt-tolerant haplotype
41 圣豆17 Shengdou 17 0.31 4 4 3 敏感单倍型 Salt-sensitive haplotype
42 中黄302 Zhonghuang 302 0.30 4 3 1 耐盐单倍型 Salt-tolerant haplotype
43 中黄318 Zhonghuang 318 0.30 4 1 2 耐盐单倍型 Salt-tolerant haplotype
44 齐转19147 Qizhuan 19147 0.29 4 5 3 敏感单倍型 Salt-sensitive haplotype
45 齐转20364 Qizhuan 20364 0.27 4 5 5 杂合 Heterozygous haplotype
46 中黄616 Zhonghuang 616 0.25 5 4 4 敏感单倍型 Salt-sensitive haplotype
47 齐农30 Qinong 30 0.24 5 3 5 敏感单倍型 Salt-sensitive haplotype
48 中黄618 Zhonghuang 618 0.21 5 4 4 敏感单倍型 Salt-sensitive haplotype

图1

根系分析系统识别图例 A: 豆芽黑白二值图像; B: 胚根识别分析图像。"

图2

8个大豆种质在对照和150 mmol L-1 NaCl处理条件下2 d (A)和3 d (B) 的表型"

图3

不同处理(对照和150 mmol L-1 NaCl)大豆的发芽率及芽长 A、B: 2 d发芽率和芽长; C、D: 3 d发芽率和芽长。数据为3次生物学重复均值, 误差线为标准误(n = 3)。柱子上方小写字母表示同一种质的不同处理在0.05概率水平差异显著。"

表2

不同大豆种质萌发期相对发芽率和相对芽长变异"

指标
Index name		 最小值
Minimum		 最大值
Maximum		 平均值
Average		 标准偏差
Standard deviation		 偏度
Skewness		 峰度
Kurtosis		 变异系数
Coefficient of variation
2 d相对发芽率
Relative germination rate after 2 days (%)		 72 100 92.17 0.11 -0.75 -0.98 0.12
3 d相对发芽率
Relative germination rate after 3 days (%)		 74 100 95.76 0.10 -1.31 2.30 0.10
2 d相对芽长
Relative radicle length after 2 days		 0.26 0.52 0.39 0.10 -0.18 -1.80 0.25
3 d相对芽长
Relative radicle length after 3 days		 0.21 0.61 0.44 0.12 -0.63 0.35 0.27

表3

8个种质盐处理2 d和3 d相对发芽率和相对芽长差异分析"

大豆名称
Soybean name		 2 d相对发芽率
Relative germination rate after 2 d (%)		 3 d相对发芽率
Relative germination rate after 3 d (%)		 2 d相对芽长
Relative radicle length after 2 d		 3 d相对芽长
Relative radicle length after 3 d
中吉602 Zhongji 602 79±14 a 93±9 a 0.38±0.08 a 0.55±0.10 a
中黄685 Zhonghuang 685 100±0 a 97±5 a 0.50±0.06 a 0.61±0.06 a
中黄311 Zhonghuang 311 86±8 a 100±5 a 0.30±0.06 a 0.41±0.07 abc
中黄39 Zhonghuang 39 100±6 a 100±0 a 0.52±0.04 a 0.50±0.07 ab
中黄324 Zhonghuang 324 100±8 a 90±0 a 0.44±0.03 a 0.48±0.02 ab
中黄349 Zhonghuang 349 100±0 a 100±0 a 0.46±0.07 a 0.44±0.04 ab
中黄616 Zhonghuang 616 72±7 a 100±0 a 0.26±0.04 a 0.34±0.08 bc
中黄618 Zhonghuang 618 94±12 a 74±19 a 0.26±0.07 a 0.21±0.04 c

图4

大豆萌发期、出苗期和苗期耐盐等级差异分析 A: 萌发期种质间相对芽长差异分析; B: 萌发期、出苗期和苗期耐盐种质维恩图。小写字母表示在0.05概率水平差异显著。"

表4

大豆3个发育时期耐盐等级的相关性分析"

耐盐等级
Salt tolerance
level		 萌发期
Germination stage		 出苗期
Emergence stage		 苗期
Seedling stage
萌发期Germination stage 1
出苗期Emergence stage 0.282 1
苗期Seedling stage 0.136 0.093 1
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