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作物学报 ›› 2024, Vol. 50 ›› Issue (12): 3013-3024.doi: 10.3724/SP.J.1006.2024.44060

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

生育期基因E1~E4不同突变组合对大豆纬度适应性的影响

方然1(), 袁丽媚1, 王玉林1, 芦思佳1,2, 孔凡江1,2, 刘宝辉1,2,*(), 孔令平1,2,*()   

  1. 1广州大学生命科学学院 / 分子遗传与进化创新研究中心, 广东广州 510006
    2广东省植物适应性与分子设计重点实验室, 广东广州 510006
  • 收稿日期:2024-04-12 接受日期:2024-08-15 出版日期:2024-12-12 网络出版日期:2024-08-29
  • 通讯作者: *刘宝辉, E-mail: liubh@gzhu.edu.cn; 孔令平, E-mail: lpkong@gzhu.edu.cn
  • 作者简介:E-mail: ranfang117@163.com
  • 基金资助:
    国家自然科学基金青年基金项目(31901569)

Effect of allelic combinations of soybean maturity loci E1/E2/E3/E4 on latitude adaptation

FANG Ran1(), YUAN Li-Mei1, WANG Yu-Lin1, LU Si-Jia1,2, KONG Fan-Jiang1,2, LIU Bao-Hui1,2,*(), KONG Ling-Ping1,2,*()   

  1. 1College of Life Sciences, Guangzhou University / Innovative Research Center of Molecular Genetics and Evolution, Guangzhou 510006, Guangdong, China
    2Guangdong Key Laboratory of Plant Adaptation and Molecular Design, Guangzhou 510006, Guangdong, China
  • Received:2024-04-12 Accepted:2024-08-15 Published:2024-12-12 Published online:2024-08-29
  • Contact: *E-mail: liubh@gzhu.edu.cn; E-mail: lpkong@gzhu.edu.cn
  • Supported by:
    Youth Found of National Natural Science Foundation of China(31901569)

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摘要：

大豆作为重要的油料作物, 是人类优质蛋白和食用油的主要来源之一。大豆产量、种子品质和生育期性状密切相关, 生育期性状主要受一系列生育期相关基因的控制。本研究以Harosoy为遗传背景创制了E1~E4不同基因组合的16种近等基因系材料, 种植在河北石家庄和安徽合肥试验田, 调查了生育期、品质及产量性状, 以此来了解E1~E4不同突变组合对中纬度种植区域的适应性。研究结果表明, 16种近等基因系材料, 对光周期敏感程度不同, 开花期存在差异, WT和e4单突近等基因系由于晚花晚熟、产量低, 不适合在石家庄种植, 而所有的近等基因系在合肥种植时均能正常成熟。E1~E4不同等位基因组合还影响株高、节间距、单株产量、品质等农艺性状。本研究发现, 在长日照条件下, e3或e4单独突变可以提前开花, 同时还会使大豆产生避荫反应, 株高增高、节间距变大。测量种子的蛋白质、油分和蔗糖含量时发现, WT材料在较高纬度的石家庄地区种植时, 种子不能正常成熟, 含油量最低, 而蔗糖的含量最高。整体来看, 其余近等基因系的种子在石家庄地区种植时总油分、蔗糖含量高于合肥, 而蛋白含量总体低于合肥地区。因此, 在评估大豆品种的纬度适应性时, 需综合考察生育期基因对光周期敏感性、品质和产量的影响。

关键词: 大豆, 生育期, 产量, 品质, 适应性

Abstract:

Soybean, as an important oil crop, is one of the main sources of high-quality protein and edible oil. Soybean yield and seed quality are closely related to growth-period traits, which are mainly controlled by a series of genes associated with the growth period. In this study, 16 near-isogenic lines (NILs) of E1-E4 were developed using Harosoy as the genetic background and were planted in experimental fields in Shijiazhuang and Hefei. The growth period, seed quality, and yield traits were investigated to understand the adaptability of different combinations of E1-E4 mutants to mid-latitude planting areas. The results showed that the 16 NILs had different photoperiod sensitivities and flowering times. WT and e4 NILs were unsuitable for planting in Shijiazhuang due to late flowering and low yield, while all NILs matured normally when planted in Hefei. Different allelic combinations of E1-E4 also affected plant height, node length, yield per plant, and seed quality. We found that e3 or e4 mutations could lead to early flowering under long-day conditions and simultaneously induce a shading response, resulting in taller plants and longer node lengths. We measured the protein, oil, and sucrose content of the seeds and found that the seeds of WT could not mature normally, exhibiting the lowest oil content and the highest sucrose content. Overall, seeds from the other NILs, when planted in Shijiazhuang, showed higher oil and sucrose content compared to those planted in Hefei but lower protein content. Therefore, to evaluate the latitude adaptability of soybean cultivars, it is necessary to comprehensively examine the effects of growth-period genes on photoperiod sensitivity, seed quality, and yield.

Key words: soybean, growth period, yield, seed quality, adaptability

表1

以Harosoy为遗传背景的E1~E4近等基因系材料"

编号
Number		 基因型
Genotype		 类型
Type		 编号
Number		 基因型
Genotype		 类型
Type
1 E1, E2, E3, E4 野生型 Wild type (WT) 9 e1, E2, e3, E4 e1e3
2 E1, e2, E3, E4 e2 10 e1, E2, E3, e4 e1e4
3 E1, E2, e3, E4 e3 11 e1, e2, E3, E4 e1e2
4 E1, E2, E3, e4 e4 12 e1, e2, e3, E4 e1e2e3
5 e1, E2, E3, E4 e1 13 E1, e2, e3, e4 e2e3e4
6 E1, e2, e3, E4 e2e3 14 e1, e2, E3, e4 e1e2e4
7 E1, e2, E3, e4 e2e4 15 e1, E2, e3, e4 e1e3e4
8 E1, E2, e3, e4 e3e4 16 e1, e2, e3, e4 e1e2e3e4

表2

近等基因系E1~E4基因型的鉴定引物"

基因
Gene		 变异类型
Variation type		 引物序列
Primer sequence (5°-3°)		 标记方法
Marker type		 限制性内切酶
Restriction enzyme		 参考文献
Reference
E1 e1 E1-NS: AACACTCAAAACACTCAAATTAAGCC
E1-RV: TCCGATCTCATCACCTTTCC		 dCAPS Hinf I [8]
E2 e2 E2-dcap-fw: GAAGCCCATCAGAGGCATGTCTTATT
E2-dcap-rv: GAGGCAGAGCCAAAGCCTAT		 dCAPS Dra I [10,31]
E3 e3 E3_08557FW: TGGAGGGTATTGGATGATGC
E3Ha_1000RV: CGGTCAAGAGCCAACATGAG
E3T_0716RV: GTCCTATACAATTCTTTACGACG		 FLP [32]
E4 e4 PhyA2-For (PhyATHF3): AGACGTAGTGCTAGGGCTAT
PhyA2-Rev/E4 (1260R): GCATCTCGCATCACCAGATCA
PhyA2-Rev/e4 (TSR): GCTCATCCCTTCGAATTCAG		 FLP [12]

图1

Harosoy近等基因系材料E1~E4基因型鉴定结果 A~D分别是表1中16种近等基因系材料的E1~E4基因分型结果; M: marker, 2000 bp DNA ladder。"

图2

近等基因系材料R1期统计分析 A, B分别代表石家庄和合肥两地的R1期统计及分析结果。图中与野生型数据比较的方差分析结果, *、**、***分别表示在0.05、0.01、0.001水平差异显著。DAE: 开花后天数(天)。"

图3

近等基因系节数和株高结果统计 A, B为石家庄田间材料节数和株高的数据及分析; C, D为合肥试验田的结果。图中与野生型数据比较的方差分析结果, *、**、***分别表示在0.05、0.01、0.001概率水平差异显著。"

图4

近等基因系材料的单株粒重比较 A, B分别代表近等基因系在石家庄和合肥种植时的单株粒重统计及分析结果。图中与野生型数据比较的方差分析结果, *、**、***分别表示在0.05、0.01、0.001概率水平差异显著。"

图5

近等基因系中野生型与e1/e2/e3/e4四突变体植株生长状态 A, B: 种植在石家庄实验田的近等基因系中野生型与e1e2e3e4四突变体收获时植株形态; C, D: 种植在合肥实验田收获时植株形态。图中标尺为5 cm。"

图6

不同光周期条件下e3/E4和 e3/e4突变体产量相关性状统计 A~C: 以Willams_82作为遗传背景, 利用CRISPR-Cas9技术创制的e3/E4和e3/e4突变体, 在光周期为20 h光照/4 h黑暗的培养箱种植, 成熟后的株高、节数、平均节间距、分枝数、单株粒数、单株粒重数据统计与分析以及植株形态展示; A表示e3/E4和e3/e4之间比较的方差分析结果, P值通过单因素方差分析(One-way ANOVA)获得; D~E: Willams_82及e3/e4突变体, 种植在16 h光照/8 h黑暗的培养箱中, 开花后株高、节数、节间距产量相关数据的统计与分析以及植株形态展示; D表示与野生型数据比较的方差分析结果, P值通过单因素方差分析(One-way ANOVA)获得。"

图7

近等基因系材料的种子品质性状比较 A~C代表在石家庄收获的近等基因系种子, 蛋白、油分和蔗糖含量品质性状统计及分析结果; D~F代表在合肥收获的种子品质性状统计及分析结果。图中与野生型数据比较的方差分析结果, *、**、***分别表示在0.05、0.01、0.001概率水平差异显著。"

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