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作物学报 ›› 2019, Vol. 45 ›› Issue (11): 1628-1637.doi: 10.3724/SP.J.1006.2019.82064

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

籼稻背景下导入Wx in等位基因改良稻米食味和理化品质

杨勇1,陆彦1,2,郭淑青1,石仲慧1,赵杰1,范晓磊1,李钱峰1,刘巧泉1,*(),张昌泉1,*()   

  1. 1 扬州大学农学院 / 植物功能基因组学教育部重点实验室 / 江苏省作物基因组学和分子育种重点实验室 / 粮食作物现代产业技术协同创新中心, 江苏扬州 225009
    2 扬州大学测试中心, 江苏扬州 225009
  • 收稿日期:2018-12-14 接受日期:2019-06-20 出版日期:2019-11-12 网络出版日期:2019-07-15
  • 通讯作者: 刘巧泉,张昌泉
  • 作者简介:E-mail: 1046354026@qq.com
  • 基金资助:
    本研究由国家重点研发计划项目(2016YFD0100501);国家自然科学基金项目(31872860);国家自然科学基金项目(31561143008);江苏省科技计划项目(BE2018357);江苏省科技计划项目(BK20160464);江苏省高等学校自然科学研究项目(16KJB210011);杂交水稻国家重点实验室(湖南杂交水稻研究中心)开放课题(2018KF04Hunan Hybrid Rice Research Center);扬州大学农学院农学专业本科生创新训练计划项目资助

Improvement of rice eating quality and physicochemical properties by introgression of Wx in allele in indica varieties

YANG Yong1,LU Yan1,2,GUO Shu-Qing1,SHI Zhong-Hui1,ZHAO Jie1,FAN Xiao-Lei1,LI Qian-Feng1,LIU Qiao-Quan1,*(),ZHANG Chang-Quan1,*()   

  1. 1 Jiangsu Key Laboratory for Crop Genomics and Molecular Breeding / Key Laboratory of Plant Functional Genomics of Ministry of Education / Co-Innovation Center for Modern Production Technology of Grain Crops, Agricultural College of Yangzhou University, Yangzhou 225009, Jiangsu, China
    2 Instrumental Analysis Center, Yangzhou University, Yangzhou 225009, Jiangsu, China
  • Received:2018-12-14 Accepted:2019-06-20 Published:2019-11-12 Published online:2019-07-15
  • Contact: Qiao-Quan LIU,Chang-Quan ZHANG
  • Supported by:
    This study was supported by the National Key Research and Development Program of China(2016YFD0100501);the National Natural Science Foundation of China(31872860);the National Natural Science Foundation of China(31561143008);the Government of Jiangsu Province(BE2018357);the Government of Jiangsu Province(BK20160464);the Natural Science Foundation of the Jiangsu Higher Education Institutions of China(16KJB210011);the Open Research Fund of State Key Laboratory of Hybrid Rice(Hunan Hybrid Rice Research Center)(2018KF04Hunan Hybrid Rice Research Center);the Personnel Training Program for Undergraduates in Agricultural College of Yangzhou University

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摘要：

水稻Wx b等位基因已广泛用于籼稻的品质改良, 但携带该等位基因的一些籼稻米饭往往偏软, 仍需进一步改良。为明确籼稻背景下导入Wx in等位基因对稻米食味品质和理化品质的效应, 分别以携带Wx in的IR64和携带Wx b的9311为供体, 以携带Wx a的籼稻SIR3611 (3611)为受体, 基于分子标记辅助选择, 通过杂交和连续回交的方式构建了3611背景下携带Wx in和Wx b的近等基因系。系统比较了不同近等基因系间的农艺性状以及稻米的食味和理化品质。结果表明, 近等基因系与受体亲本3611的主要农艺性状基本接近, 无显著差异。NIL(Wx in)稻米的表观直链淀粉含量较亲本3611极显著下降而胶稠度极显著增加。NIL(Wx b)稻米表观直链淀粉含量最低且与之对应的胶稠度最高。近等基因系NIL(Wx in)和NIL(Wx b)稻米的食味值较亲本极显著提高。NIL(Wx in)和NIL(Wx b)稻米的GBSSI丰度与对应的表观直链淀粉含量具有明显的正相关。稻米粉的黏滞性谱、热糊化特性和晶体结构与直链淀粉含量显著相关性。本研究为在我国籼稻品种品质改良中有效利用Wx in等位基因提供了重要依据。

关键词: 水稻, 食味品质, Wx等位基因, 表观直链淀粉含量, 分子标记辅助选择

Abstract:

Nowadays, the Wx b allele has been widely used to improve grain quality of indica rice. However, some indica varieties carrying Wx b allele usually has a much softer texture, which is not favored by consumers in South China. So the grain quality of these varieties needs to be further improved. To understand the effect of Wx in allele on rice eating quality and physicochemical properties in indica rice, we developed two Near-Isogenic Lines (NILs) carrying Wx in and Wx b alleles by crossing an indica variety 3611 (receptor, carrying Wx a) with IR64 (carrying Wx in) and 9311 (carrying Wx b), and seven times of backcrossing based on molecular marker assistant selection (MAS). The Wx effects in controlling the synthesis of amylose, grain quality, and physicochemical properties were investigated. There were non-significant differences in the agronomic traits among the NILs. However, for grain quality characters, we found that the NIL(Wx in) rice showed significantly lower apparent amylose content (AAC) and higher gel consistency (GC), compared with the wild type 3611. Besides, the NIL(Wx b) rice showed the lowest AAC and highest GC among three lines. The NIL(Wx in) rice had a significantly higher taste value than the wild type 3611, while the NIL(Wx b) rice exhibited the highest taste value among the three samples. The granule-bound starch synthase I (GBSSI) level was the highest in 3611, moderate in NIL(Wx in) and lowest in NIL(Wx b), which showed a positive correlation with the AAC level. Also, the starch viscosity, thermal gelatinization property and crystal structure of different rice flours had a high correlation with the AAC level. To sum up, our results proved that both Wx in and Wx b allele can improve the grain quality in 3611 background, and what is more, the Wx in allele might be more useful for the improvement of grain quality in indica rice.

Key words: Oryza sativa L., eating quality, Wx allele, apparent amylose content, molecular marker assisted selection

图1

近等基因系构建过程(A)与近等基因系糙米表型(B) MAS: 分子标记辅助选择; NIL: 近等基因系。"

图2

不同Wx等位基因特异分子标记的检测 A图为利用QRM190分子标记区分Wxa和Wxb的PCR检测结果, 泳道1为3611, 2为9311, 3~7为近等基因系NIL(Wxb); B图为利用基因特异分子标记区分Wxa和Wxin的PCR检测结果, 泳道1为3611, 2为IR64, 3~7为近等基因系NIL(Wxin)。"

表1

近等基因系主要农艺性状表现"

品系
Line		 株高
Plant height (cm)		 有效分蘖
Tiller number		 主穗长
Main panicle length (cm)		 结实率
Seed setting rate (%)		 千粒重
1000-kernel weight (g)
3611(Wxa) 117.8±1.2 a 5.6±0.9 a 26.5±0.4 a 91.2±2.2 a 30.7±0.8 a
NIL(Wxin) 118.2±2.3 a 5.2±0.8 a 26.7±0.6 a 90.5±3.1 a 29.8±0.5 a
NIL(Wxb) 115.0±1.5 a 6.0±0.6 a 25.8±0.4 a 91.5±2.8 a 31.0±0.2 a

图3

近等基因系成熟种子中GBSS I蛋白游离态(A)和结合态(B)的SDS-PAGE分析 M: 蛋白质分子质量标准; 1~2: 3611(Wxa); 3~4: NIL(Wxin); 5~6: NIL(Wxb); 箭头所示为60 kD的GBSSI蛋白。"

表2

不同近等基因系稻米理化和外观品质"

品系
Line		 表观直链淀粉含量
Apparent amylose content (%)		 胶稠度
Gel consistency
(mm)		 食味值
Taste value		 垩白粒率
Chalkiness rate
(%)		 垩白度
Chalkness degree
(%)
3611(Wxa) 25.51±1.48 a 42.35±4.17 c 43.15±1.35 c 18.62±0.87 a 2.89±1.05 a
NIL(Wxin) 19.85±0.27 b 78.54±2.32 b 55.09±2.51 b 13.04±0.30 b 1.93±0.06 c
NIL(Wxb) 14.78±0.55 c 87.62±3.48 a 60.51±1.87 a 13.05±0.40 b 2.03±0.32 b

图4

不同近等基因系稻米粉RVA谱分析"

表3

不同近等基因系稻米RVA特征值"

品系
Line		 峰值黏度
Peak viscosity (cP)		 热浆黏度
Hot paste
viscosity (cP)		 崩解值
Breakdown
(cP)		 冷胶黏度
Cool paste
viscosity (cP)		 回复值
Setback
(cP)		 峰值时间
Peak time
(min)		 起浆温度
Peak temperature (℃)
3611(Wxa) 3143.2±35.2 b 2607.4±27.1 a 536.4±12.8 c 3960.2±31.6 a 817.6±18.6 a 6.4±0.1 a 70.80±1.1 b
NIL(Wxin) 2961.5±47.1 c 2211.5±31.1 c 750.6±9.3 b 3503.4±28.7 b 514.8±12.6 b 6.3±0.1 a 71.65±1.0 b
NIL(Wxb) 3628.5±28.3 a 2383.3±30.4 b 1245.7±10.5 a 3390.3±21.5 c -238.7±9.5 c 6.4±0.1 a 73.25±1.0 a

图5

不同近等基因系米粉的DSC分析"

表4

不同近等基因系稻米米粉热糊化参数"

品系
Line		 起始温度
T0 (°C)		 峰值温度
Tp (°C)		 终止温度
Tc (°C)		 热焓值
ΔH (J G-1)
3611(Wxa) 62.89±0.21 c 67.68±0.62 c 76.54±0.31 c 6.39±0.30 c
NIL(Wxin) 63.57±0.12 b 69.53±0.70 b 77.02±0.07 b 8.01±0.10 b
NIL(Wxb) 64.86±0.41 a 71.33±0.60 a 77.91±0.46 a 8.53±0.11 a

图6

不同近等基因系米粉的X射线衍射(A)和红外光谱分析(B)"

表5

不同近等基因系米粉的X射线衍射和傅里叶变换红外光谱参数"

品系
Line		 结晶度
Crystallinity (%)		 峰强比值
IR ratio of 1045/1022 cm-1
3611(Wxa) 27.35±0.31 c 0.64±0.01 c
NIL(Wxin) 29.17±0.28 b 0.76±0.01 b
NIL(Wxb) 32.04±0.41 a 0.80±0.01 a
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