Effects of Different Combinations of Wx and ALK Main Alleles on Rice Grain Quality

doi:10.16819/j.1001-7216.2020.9140

Abstract

Abstract:

【Objective】Rice eating and cooking qualities are mainly controlled by two key genes, Wx and ALK. There are multiple alleles of both genes among rice cultivars. In this study, our aim is to clarify the effects of the main allelic variation combinations of Wx and ALK genes on rice grain quality.【Method】Three main alleles of Wx (Wx^a, Wx^b and wx) and two of ALK (ALK^a and ALK^c) were combined within a series of near-isogenic lines (NILs) in the japonica Nipponbare (Nip) background. The effects of different allelic combinations on grain quality were determined by measuring the physical and chemical characteristics of NILs.【Result】Gelatinization temperature (GT) was regulated by the major gene ALK and the minor gene Wx. Under the same ALK allele background, there was no significant difference in GT between rice lines carrying wx and Wx^b alleles, but the GT of Nip-Wx^a rice lines was significantly lower than that of Nip-Wx^b and Nip-wx. The starch viscosity was greatly affected by the combination of Wx and ALK alleles. The breakdown value of Nip-Wx^a rice line was lower and the setback value was higher (lower tasting value) than those of Nip-Wx^b and Nip-wx. Under the same Wx allele background, ALK allelic variation mainly affected the pasting temperature of RVA curve, but had little effect on breakdown value and setback value. Nip-Wx^a/ALK^a and Nip-Wx^a/ALK^c rice grains had poor tasting value due to the high AC and breakdown value and low GC. But the protein content in above two NILs was higher than other rice lines. Nip-wx/ALK^a and Nip-Wx^b/ALk^a rice grains had better tasting value of hot cooked and cooled cooked rice due to the moderate AC, GC and protein content, as well as lower GT. Although the GT of Nip-Wx^b/ALK^c grains was higher, the moderate AC and high GC of this line resulted in a slight decrease in the tasting value of cooled cooked rice compared with hot cooked rice. The tasting value of hot cooked rice of Nip-wx/ALK^c rice remained the same as those of Nip-wx/ALK^a, Nip-Wx^b/ALK^a and Nip-Wx^b/ALK^c, but the tasting value of cooled rice decreased dramatically because of its high GT. 【Conclusion】Amylose content (AC) and gel consistency (GC) were mainly controlled by Wx gene; while ALK gene had no significant effect on AC, but a slight effect on GC.

Key words: waxy gene, alkali digestion gene, allelic variation, near isogenic lines, grain quality

摘要：

【目的】稻米蒸煮和食味品质主要受Wx和ALK两个主效基因的调控。这两个基因在栽培稻中存在多个复等位变异,本研究旨在明确Wx和ALK基因的主要等位变异及其组合对稻米品质的影响。【方法】将Wx的3个主要等位基因（Wx^a、Wx^b和wx）与ALK的2个主要等位基因（ALK^a和ALK^c）进行不同的组合,在粳稻品种日本晴（Nip）背景下创建6个含有Wx和ALK基因不同等位变异组合的近等基因系。通过测定近等基因系的稻米理化品质明确不同等位基因及其组合对稻米品质的效应。【结果】在相同的ALK基因型背景下,含Wx^a等位基因稻米的糊化温度极显著低于wx型和Wx^b型的近等基因系。稻米黏滞性受Wx和ALK等位基因组合影响较大,Wx等位变异主要影响稻米的峰值黏度、峰值时间和冷胶黏度;在相同Wx背景下,ALK等位基因主要影响稻米RVA谱的起浆温度,对稻米崩解值和消减值的影响不显著。总体来看,Nip-Wx^a/ALK^a和Nip-Wx^a/ALK^c型稻米具有较高直链淀粉含量和崩解值,较低的胶稠度,因而食味值较差;Nip-wx/ALK^a和Nip-Wx^b/ALK^a型水稻具有适中的直链淀粉含量、较软的胶稠度以及较低的糊化温度,因此这两个近等基因系热饭和冷饭的食味值较好;虽然Nip-Wx^b/ALK^c的糊化温度较高,但因其适中的直链淀粉含量和较高的胶稠度,其冷饭食味值降幅较小;Nip-wx/ALK^c稻米的热饭食味值与Nip-wx/ALK^a、Nip-Wx^b/ALK^a和Nip-Wx^b/ALK^c无差异,但因其较高的糊化温度,其冷饭食味值显著下降。【结论】Wx基因变异主要影响稻米直链淀粉含量和胶稠度,对糊化温度有一定的影响;ALK基因变异主要影响糊化温度,对直链淀粉含量无显著影响,对胶稠度存在微效影响。

关键词: 蜡质基因, 糊化温度基因, 等位基因, 近等基因系, 稻米品质

CLC Number:

Zhuanzhuan CHEN, Yong YANG, Linhao FENG, Ye SUN, Changquan ZHANG, Xiaolei FAN, Qianfeng LI, Qiaoquan LIU. Effects of Different Combinations of Wx and ALK Main Alleles on Rice Grain Quality[J]. Chinese Journal OF Rice Science, 2020, 34(3): 228-236.

陈专专, 杨勇, 冯琳皓, 孙晔, 张昌泉, 范晓磊, 李钱峰, 刘巧泉. Wx与ALK主要等位基因不同组合对稻米品质的影响[J]. 中国水稻科学, 2020, 34(3): 228-236.

Figures/Tables 8

References 38

[1]	Vandeputte G E, Delcour J A.From sucrose to starch granule to starch physical behaviour: A focus on rice starch[J]. Carbohydrate Polymers, 2004, 58(3): 245-266.
[2]	Singh N, Kaur L, Sandhu K S, Kaur J, Nishinari K.Relationships between physicochemical, morphological, thermal, rheological properties of rice starches[J]. Food Hydrocolloid, 2006, 20(4): 532-542.
[3]	包劲松. 稻米淀粉品质遗传与改良研究进展[J]. 分子植物育种, 2007, 5(6): 1-20.
	Bao J S.Progress in sutdies on inheritance and improvement of rice starch qualiy[J]. Molecular Plant Breeding, 2007, 5(6): 1-20. (in Chinese with English abstract)
[4]	Wang Z Y, Zheng F Q, Shen G Z, Gao J P, Snustad D P, Li M G, Zhang J L, Hong M M.The amylose content in rice endosperm is related to the post-transcriptional regulation of the waxy gene[J]. Plant Journal, 1995, 7(4): 613-622.
[5]	Tian Z X, Qian Q, Liu Q Q, Yan M X, Liu X F, Yan C J, Liu G F, Gao Z Y, Tang S Z, Zeng D L, Wang Y H, Yu J M, Gu M H, Li J Y.Allelic diversities in rice starch biosynthesis lead to a diverse array of rice eating and cooking qualities[J]. Proceedings of the National Academy of Sciences USA, 2009, 106(51): 21760-21765.
[6]	Cai X L, Wang Z Y, Xing Y Y, Zhang J L, Hong M M.Aberrant splicing of intron 1 leads to the heterogeneous 5′ UTR and decreased expression of waxy gene in rice cultivars of intermediate amylose content[J]. Plant Journal, 1998, 14(4): 459-465.
[7]	Gao Z Y, Zeng D L, Cheng F M, Tian Z X, Guo L B, Su Y, Yan M X, Jiang H, Dong G J, Huang Y C, Han B, Li J Y, Qian Q.ALK, the key gene for gelatinization temperature, is a modifier gene for gel consistency in rice[J]. Journal of Integrative Plant Biology, 2011, 53(9): 756-765.
[8]	Zhang C Q, Zhu J H, Chen S J, Fan X L, Lu Y, Wang M, Yu H X, Yi C D, Tang S Z, Gu M H, Liu Q Q.Wxlv, the ancestral allele of rice Waxy gene[J]. Molecular Plant, 2019, 12(8): 1157-1166.
[9]	朱霁晖, 张昌泉, 顾铭洪, 刘巧泉. 栽培稻中Wx基因的等位变异及育种利用研究进展[J]. 中国水稻科学, 2015, 29(4): 431-438.
	Zhu J H, Zhang C Q, Gu M H, Liu Q Q.Progress in the allelic variation of Wx gene and its application in rice[J]. Chinese Journal of Rice Science, 2015, 29(4): 431-438. (in Chinese with English abstract)
[10]	Sano Y.Differential regulation of waxy gene expression in rice endosperm[J]. Theoretical and Applied Genetics, 1984, 68(5): 467-473.
[11]	范名宇, 王晓菁, 王旭虹, 唐亮, 徐铨, 徐正进. 稻米支链淀粉结构的研究进展[J]. 中国水稻科学, 2017, 31(2): 124-132.
	Fan M Y, Wang X J, Wang X H, Tang L, Xu Q, Xu Z J.Research advances of amylopectin structure in rice[J]. Chinese Journal of Rice Science, 2017, 31(2): 124-132. (in Chinese with English abstract)
[12]	Jeon J S, Ryoo N, Hahn T R, Walia H, Nakamura Y.Starch biosynthesis in cereal endosperm[J]. Plant Physiology and Biochemistry, 2010, 48(6): 383-392.
[13]	孙业盈, 吕彦, 董春林, 王平荣, 黄晓群, 邓晓建. 水稻Wx基因与稻米AC、GC和GT的遗传关系[J]. 作物学报, 2005, 31(5): 535-539.
	Sun Y Y, Lv Y, Dong C L, Wang P R, Hang X Q, Deng X J.Genetic relationship among Wx gene, AC, GC and GT of rice[J]. Acta Agronomica Sinica, 2005, 31(5): 535-539. (in Chinese with English abstract)
[14]	刘玉花, 栾丽, 龙文波, 王兴, 孔繁伦, 何涛, 涂升斌. 同源四倍体和二倍体水稻Wx基因与淀粉品质的遗传关系[J]. 中国水稻科学, 2007, 21(2): 143-149.
	Liu Y H, Luan L, Long W B, Wang X, Kong F L, He T, Tu S B.Genetic relationships between Wx gene and starch quality in autotereaploid and diploid rice[J]. Chinese Journal of Rice Science, 2007, 21(2): 143-149. (in Chinese with English abstract)
[15]	Varavinit S, Shobsngob S, Varanyanond W, Chinachoti P, Naivikul O.Effect of amylose content on gelatinization, retrogradation and pasting properties of flours from different cultivars of Thai rice[J]. Starch-Stärke, 2003, 55(9): 410-415.
[16]	苏文丽, 向珣朝, 徐艳芳, 龙小林, 康翠芳, 许顺菊. 水稻SSIIa基因型对成糊温度校准的影响及糊化温度与直链淀粉含量的关系分析[J]. 植物遗传资源学报, 2014, 15(2): 348-353.
	Su W L, Xiamg X C, Xu Y F, Long X L, Kang C F, Xu S J.The influence of SSIIa genotype on the calibration of pasting temperature of rice and the relationship between gelatinization temperature and amylose content[J]. Journal of Plant Genetic Resources, 2014, 15(2): 348-353. (in Chinese with English abstract)
[17]	Xiang X C, Kang C F, Xu S J, Yang B W.Combined effects of Wx and SSIIa haplotypes on rice starch physicochemical properties[J]. Journal of the Science of Food and Agriculture, 2016, 97(4): 1229-1234.
[18]	Murray M G, Thompson W F.Rapid isolation of high molecular weight plant DNA[J]. Nucleic Acids Research, 1980, 8(19): 4321-4325.
[19]	Fan X L, Li Y Q, Lu Y, Zhang C Q, Li E P, Li Q F, Tao K Y, Yu W W, Wang J D, Chen Z Z, Liu Q Q.The interaction between amylose and amylopectin synthesis in rice endosperm grown at high temperature[J/OL].Food Chemistry, 2019, 301: 125258.
[20]	张昌泉, 潘立旭, 周兴忠, 李钱峰, 刘巧泉. 不同Wx等位组合对杂交稻天优3611品质的影响[EB/OL]. 北京:中国科技论文在线[2017-04-28]. .
	Zhang C Q, Pan L X, Zhou X Z, Li Q F, Liu Q Q. The combined effects of different Wx allele on grain quality of hybrid rice Tianyou 3611[EB/OL]. Beijing: Chinese Science Paper Online [2017-04-28]. ((in Chinese with English abstract)
[21]	Zhang C Q, Zhou L H, Zhu Z B, Lu H W, Zhou X Z, Qian Y T, Li Q F, Lu Y, Gu M H, Liu Q Q.Characterization of grain quality and starch fine structure of two japonica rice (Oryza sativa) cultivars with good sensory properties at different temperatures during the filling stage[J]. Journal of Agricultural and Food Chemistry, 2016, 64(20): 4048-4057.
[22]	Zhang C Q, Zhu L J, Shao K, Gu M H, Liu Q Q.Toward underlying reasons for rice starches having low viscosity and high amylose: Physiochemical and structural characteristics[J]. Journal of the Science of Food and Agriculture, 2013, 93(7): 1543-1551.
[23]	Zhu L J, Liu Q Q, Sang Y, Gu M H, Shi Y C.Underlying reasons for waxy rice flours having different pasting properties[J]. Food Chemistry, 2010, 120(1): 94-100.
[24]	戴宏林, 吴小骏. 用凯氏定氮法测定植物干样品中的氮含量[J]. 扬州大学学报: 农业与生命科学版, 1995, 16(3): 70.
	Dai H L, Wu X J.Determination of nitrogen in dry plant samples by Kjeldahl nitrogen determination[J]. Journal of Yangzhou University, 16(3): 70. (in Chinese with English abstract)
[25]	马玉清, 李仕贵, 王玉平, 黎汉云, 高克铭. 籼型杂交水稻稻米品质的初步研究[J]. 西南农业学报, 2002, 15(1): 28-31.
	Ma Y Q, Li S G, Wang Y P, Li H Y, Gao K M.Preliminary study on quality traits of indica hybrid rice[J]. Southwest China Journal of Agricultural Science, 2002, 15(1): 28-31. (in Chinese with English abstract)
[26]	Bao J S, Sun M, Zhu L H, Corke H.Analysis of quantitative trait loci for some starch properties of rice (Oryza sativa L.): Thermal properties, gel texture and swelling volume[J]. Journal of Cereal Science, 2004, 39(3): 379-385.
[27]	夏明元, 李进波, 张建华, 万丙良, 戚华雄. 利用分子标记辅助选择技术选育具有中等直链淀粉含量的早稻品种[J]. 华中农业大学学报, 2004, 23(2): 183-186.
	Xia M Y, Li J B, Zhang J H, Wan B L, Qi H X.Breeding of early indica rice with intermediate amylose content by molecular marker-aided selection[J]. Journal of Huangzhong Agricultural University, 2004, 23(2): 183-186. (in Chinese with English abstract)
[28]	舒庆尧, 吴殿星, 夏英武, 高明尉, McClung A. 稻米淀粉RVA谱特征与食用品质的关系[J]. 中国农业科学, 1998, 31(3): 25-29.
	Shu Q Y, Wu D X, Xia Y W, Gao M W, McClung A. Relationship between RVA character and eating quality in Oryza sativa L[J]. Scientia Agricultura Sinica, 1998, 31(3): 25-29. (in Chinese with English abstract)
[29]	曾瑞珍, 张泽民, 何风华, 席章营, Talukdar A, 施军琼, 秦利军, 黄朝锋, 张桂权. 水稻Wx复等位基因的鉴定及单片段代换系的建立[J]. 中国水稻科学, 2005, 19(6): 495-500.
	Zeng R Z, Zhang Z M, He F H, Xi Z Y, Akshay T, Shi J Q, Qin L J, Huang C F, Zhang G Q.Identification of multiple alleles at the Wx locus and development of single segment substitution lines in rice[J]. Chinese Journal of Rice Science, 2005, 19(6): 495-500. (in Chinese with English abstract)
[30]	陈专专, 李先锋, 仲敏, 葛家奇, 范晓磊, 张昌泉, 刘巧泉. 籼稻背景下抑制不同ALK等位基因表达对稻米品质的影响[J]. 中国水稻科学, 2019, 33(6): 513-522.
	Chen Z Z, Li X F, Zhong M, Ge J Q, Fan X L, Zhang C Q, Liu Q Q.Effects on grain quality by down-regulation of the expression of different ALK allele in indica rice (Oryza sativa L.)[J]. Chinese Journal of Rice Science, 2019, 33(6): 513-522. (in Chinese with English abstract)
[31]	李欣, 顾铭洪, 潘学彪. 常见水稻品种稻米品质的研究[J]. 扬州大学学报:农业与生命科学版, 1987, 8(1): 1-8.
	Li X, Gu M H, Pan X B.A study on grain quality of rice for commercial varieties[J]. Journal of Yangzhou University, 1987, 8(1): 1-8. (in Chinese with English abstract)
[32]	Han X Z, Hamaker B R.Amylopectin fine structure and rice starch paste breakdown[J]. Journal of Cereal Science, 2001, 34(3): 279-284.
[33]	Ong M H, Blanshard J M V. Texture determinants of cooked, parboiled rice. II: Physicochemical properties and leaching behaviour of rice[J]. Journal of Cereal Science, 1995, 21(3): 261-269.
[34]	Wang K, Hasjim J, Wu A C, Li E P, Henry R J, Gilbert R G.Roles of GBSSI and SSIIa in determining amylose fine structure[J]. Carbohydrate Polymers, 2015, 127: 264-274.
[35]	万映秀, 邓其明, 王世全, 刘明伟, 周华强, 李平. 水稻Wx基因的遗传多态性及其与主要米质指标的相关性分析[J]. 中国水稻科学, 2006, 20(6): 603-609.
	Wan Y X, Deng Q M, Wang S Q, Liu M W, Zhou H Q, Li P.Genetic polymorphism of Wx gene and its correlation with major grain quality traits in rice[J]. Chinese Journal of Rice Science, 2006, 20(6): 603-609. (in Chinese with English abstract)
[36]	Bao J S, Corke H, He P, Zhu L H.Analysis of quantitative trait loci for starch properties of rice based on an RIL polulation[J]. Acta Botanica Sinica, 2003, 45(8): 986-994.
[37]	朱昌兰, 江玲, 张文伟, 王春明, 翟虎渠, 万建民. 稻米直链淀粉含量和胶稠度对高温耐性的QTL分析[J]. 中国水稻科学, 2006, 20(3): 248-252.
	Zhu C L, Jiang L, Zhang W W, Wang C M, Zhai H Q, Wan J M.Identifying QTLs for thermo-tolerance of amylose content and gel consistency in rice[J]. Chinese Journal of Rice Science, 2006, 20(3): 248-252. (in Chinese with English abstract)
[38]	盛文涛, 吴俊, 姚栋萍, 庄文, 邓启云. 稻米食味品质的影响因素及中国籼稻食味品质改良存在的问题与策略[J]. 杂交水稻, 2014, 29(3): 1-5.
	Sheng W T, Wu J, Yao D P, Zhuang W, Deng Q Y.Influencing factors of rice eating quality and existing problems and strategies of genetic improvement on eating quality of indica rice in China[J]. Hybrid Rice, 2014, 29(3): 1-5. (in Chinese with English abstract)

近等基因系 Near-isogenic line	Wx基因型 Wx allele	ALK基因型 ALK allele	Wx基因供体 Donor of Wx gene	ALK基因供体 Donor of ALK gene	世代 Generation
Nip-wx/ALK^a	wx	ALK^a	苏御糯 Suyunuo	日本晴 Nipponbare	BC₉F₈
Nip-wx/ALK^c	wx	ALK^c	苏御糯 Suyunuo	中恢9308 Zhonghui 9308	BC₆F₁₀
Nip-Wx^b/ALK^a	Wx^b	ALK^a	日本晴 Nipponbare	日本晴 Nipponbare
Nip-Wx^b/ALK^c	Wx^b	ALK^c	日本晴 Nipponbare	中恢9308 Zhonghui 9308	BC₈F₁₀
Nip-Wx^a/ALK^a	Wx^a	ALK^a	龙特甫 Longtefu	日本晴 Nipponbare	BC₈F₇
Nip-Wx^a/ALK^c	Wx^a	ALK^c	龙特甫 Longtefu	中恢9308 Zhonghui 9308	BC₆F₃

近等基因系 Near-isogenic line	Wx基因型 Wx allele	ALK基因型 ALK allele	Wx基因供体 Donor of Wx gene	ALK基因供体 Donor of ALK gene	世代 Generation
Nip-wx/ALK^a	wx	ALK^a	苏御糯 Suyunuo	日本晴 Nipponbare	BC₉F₈
Nip-wx/ALK^c	wx	ALK^c	苏御糯 Suyunuo	中恢9308 Zhonghui 9308	BC₆F₁₀
Nip-Wx^b/ALK^a	Wx^b	ALK^a	日本晴 Nipponbare	日本晴 Nipponbare
Nip-Wx^b/ALK^c	Wx^b	ALK^c	日本晴 Nipponbare	中恢9308 Zhonghui 9308	BC₈F₁₀
Nip-Wx^a/ALK^a	Wx^a	ALK^a	龙特甫 Longtefu	日本晴 Nipponbare	BC₈F₇
Nip-Wx^a/ALK^c	Wx^a	ALK^c	龙特甫 Longtefu	中恢9308 Zhonghui 9308	BC₆F₃

鉴定基因 Target gene	引物名称 Primer	引物序列 Primer sequence (5'-3')	区分基因型 Genotype	检测突变位置 Mutation site
Wx	QRM190-F	ATTCCTTCAGTTCTTTGTCTATCTCA	Wx^a/Wx^b	第1内含子CT序列多态性 (CT)₇ in Intron 1
	QRM190-R	TCCTGATGAACAACAGAACAACAC	Wx^a/Wx^b	第1内含子CT序列多态性 (CT)₇ in Intron 1
	ND-F	CACAGCAACAGCTAGACAACCAC	Wx/wx	第2外显子插入缺失23 bp 23 bp InDel in Exon 2
	ND-R	CACGACGACGGAGGGGAAC	Wx/wx	第2外显子插入缺失23 bp 23 bp InDel in Exon 2
ALK	4211(A/G)-F₁	GCGGGCTGAGGGACACCG	ALK^a/ALK^c	第8外显子单核苷酸多态性(A/G) A/G SNP in Exon 8
	4211(A/G)-R₁	AACGGGTCGAACGCCGACGT
	4211(A/G)-F	AACAGCAAGGTGCGCGGGTG
	4211(A/G)-R	CCACCGAGTCGCACCTCCACA

鉴定基因 Target gene	引物名称 Primer	引物序列 Primer sequence (5'-3')	区分基因型 Genotype	检测突变位置 Mutation site
Wx	QRM190-F	ATTCCTTCAGTTCTTTGTCTATCTCA	Wx^a/Wx^b	第1内含子CT序列多态性 (CT)₇ in Intron 1
	QRM190-R	TCCTGATGAACAACAGAACAACAC	Wx^a/Wx^b	第1内含子CT序列多态性 (CT)₇ in Intron 1
	ND-F	CACAGCAACAGCTAGACAACCAC	Wx/wx	第2外显子插入缺失23 bp 23 bp InDel in Exon 2
	ND-R	CACGACGACGGAGGGGAAC	Wx/wx	第2外显子插入缺失23 bp 23 bp InDel in Exon 2
ALK	4211(A/G)-F₁	GCGGGCTGAGGGACACCG	ALK^a/ALK^c	第8外显子单核苷酸多态性(A/G) A/G SNP in Exon 8
	4211(A/G)-R₁	AACGGGTCGAACGCCGACGT
	4211(A/G)-F	AACAGCAAGGTGCGCGGGTG
	4211(A/G)-R	CCACCGAGTCGCACCTCCACA

基因型 Genotype	起始糊化温度 T_o /℃	峰值糊化温度 T_p /℃	终止糊化温度 T_c /℃	热焓值 △H / (J·g^-1)
Nip-wx/ALK^a	65.7±0.2 e	72.6±0.1 d	80.5±0.1 c	9.98±0.55 bc
Nip-Wx^b/ALK^a	67.5±0.1 d	72.6±0.1 d	79.7±0.4 c	9.45±0.31 c
Nip-Wx^a/ALK^a	63.6±0.3 f	69.4±0.1 e	76.6±0.6 d	6.80±0.33 d
Nip-wx/ALK^c	75.2±0.1 b	80.6±0.0 b	86.8±0.1 a	11.93±0.35 a
Nip-Wx^b/ALK^c	76.7±0.1 a	80.8±0.0 a	86.4±0.1 a	11.03±0.29 ab
Nip-Wx^a/ALK^c	73.7±0.0 c	78.2±0.0 c	84.2±0.0 b	9.19±0.46 c