Development of Functional Markers of the Key Gene TaAGPL1 in Wheat Starch Synthesis

doi:10.13304/j.nykjdb.2024.0303

Abstract

Abstract:

Starch accounts for about 70% of wheat grain dry weight， and it is the determinant of grain weight. TaAGPL1 gene codes a large cytoplasmic subunit of adenosine diphosphate glucose pyrophosphorylase （AGPase） and plays a key role in wheat starch synthesis. From 239 modern wheat varieties approved in recent years， it was found that 13 single nucleotide polymorphism loci （SNPs） only existed on B copy of TaAGPL1 gene. After genome polymorphism analysis， they were classified into 2 haplotypes （Hap1 and Hap2）. The correlation between haplotypes and thousand grain weight was analyzed， and Hap2 was proved to be the favoured haplotype. Based on the A/C mutation at allelic mutation site 1 944 in the encoding region， a functional marker was developed for enzyme restriction amplified polymorphic sequence （CAPS）. The transcription level of TaAGPL1-1B gene and activity of AGPase between Hap1 and Hap2 varieties were measured and compared， and it was found that they were significantly increased in the grains of favoured haplotype Hap2 after grain-filling of 14 and 21 d. The research results provided theoretical basis for screening and creation of modern wheat varieties with high starch content or high thousand grain weight.

Key words: Triticum aestivum L., TaAGPL1, starch, AGPase activity, functional marker

摘要：

淀粉约占小麦籽粒干重的70%，是粒重的决定因子，TaAGPL1基因编码腺苷二磷酸葡萄糖焦磷酸化酶（AGPase）的胞质型大亚基，在小麦淀粉合成中发挥着关键作用。在近年来审定的239份小麦品种中，TaAGPL1基因序列中仅B拷贝上存在13个单核苷酸多态性位点（SNPs），根据序列多态性将小麦品种分成2种单倍型（Hap1和Hap2），将单倍型与千粒重进行相关分析发现，Hap2为优异单倍型；根据编码域1 944位点A/C碱基突变，开发了酶切扩增多态性序列（CAPS）功能标记；对Hap1和Hap2品种间TaAGPL1-1B基因转录水平和AGPase酶活性进行测定，发现它们在灌浆后14和21 d的Hap2小麦籽粒中均显著提高。研究结果为现代高淀粉含量或高千粒重小麦品种的筛选和创制提供理论基础和筛选依据。

关键词: 小麦, TaAGPL1, 淀粉, AGPase酶活性, 功能标记

CLC Number:

S512

Moming HU, Yanjun MENG, Shuyuan MIAO, Xining HU, Qing XU, Hongzhen MA, Pengfei WANG, Guoqin LIU, Guozhang KANG. Development of Functional Markers of the Key Gene TaAGPL1 in Wheat Starch Synthesis[J]. Journal of Agricultural Science and Technology, 2024, 26(11): 43-55.

胡墨明, 孟琰珺, 苗淑媛, 胡西宁, 许晴, 马红珍, 王鹏飞, 刘国芹, 康国章. 小麦淀粉合成关键基因TaAGPL1功能标记的开发[J]. 中国农业科技导报, 2024, 26(11): 43-55.

Figures/Tables 11

References 26

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基因 Gene	引物 Primer	引物序列 Primer sequence （5’-3’）	目标片段长度 Length of target sequence/bp
TaAGPL1-1A	1A-Pro-F	CGCGAGAGATCGCTCGATCG	2 796
	1A-Pro-R	ACGTTTCTGCAAATGACAGAG	2 796
	1A-CDS-F1	CATCAGTGATTTGGGCCAGGTC	1 973
	1A-CDS-R1	TCAGAAACTAGACCTCATTCAAG	1 973
	1A-CDS-F2	CATGTGGATGACAATGCTGAC	1 940
	1A-CDS-R2	TACCACTAATGCTGCATGCG	1 940
TaAGPL1-1B	1B-Pro-F	TCGCTCGCTCTTCTTCAGC	2 609
	1B-Pro-R	TGCCACCGCCGAGTATGACG	2 609
	1B-CDS-F1	ATGCTTCTCAACTTCTCGT	1 873
	1B-CDS-R1	GCTGAGACGAGTGCTAACC	1 873
	1B-CDS-F2	CATGTGGATGACAATGCTGAC	1 964
	1B-CDS-R2	CTTACTACTGATGCTGCCTGC	1 964
TaAGPL1-1D	1D-Pro-F	GGCCGGCGTCGCGAGAGCT	2 854
	1D-Pro-R	GAACATTTCTGTAAGTAACAC	2 854
	1D-CDS-F1	TACTGAGCTTTACAGAGGGCCG	2 040
	1D-CDS-R1	GACACAGTGAGGTAAGACCAG	2 040
	1D-CDS-F2	CATGTGGATGACAATGCTGAC	2 060
	1D-CDS-R2	CGGGAGTTTGTTTGCTCGCT	2 060
qTaAGPL1-1B	qTaAGPL1-F	GCCCCTGTTGGAGAGAGCCG	157
qTaAGPL1-1B	qTaAGPL1-R	TAGCAGGATCGTCTATGGCG	157
qActin	qActin-F	CCTCTCTTAGCACTTTCCAGCA	101
qActin	qActin-R	GTAAGTCCCCTTCACCGACTC	101
CAPS	CAPS-F	CATGTGCAGAAAGTGGACA	443
CAPS	CAPS-R	AGAAGGATCTGATTGTTCCA	443

基因 Gene	引物 Primer	引物序列 Primer sequence （5’-3’）	目标片段长度 Length of target sequence/bp
TaAGPL1-1A	1A-Pro-F	CGCGAGAGATCGCTCGATCG	2 796
	1A-Pro-R	ACGTTTCTGCAAATGACAGAG	2 796
	1A-CDS-F1	CATCAGTGATTTGGGCCAGGTC	1 973
	1A-CDS-R1	TCAGAAACTAGACCTCATTCAAG	1 973
	1A-CDS-F2	CATGTGGATGACAATGCTGAC	1 940
	1A-CDS-R2	TACCACTAATGCTGCATGCG	1 940
TaAGPL1-1B	1B-Pro-F	TCGCTCGCTCTTCTTCAGC	2 609
	1B-Pro-R	TGCCACCGCCGAGTATGACG	2 609
	1B-CDS-F1	ATGCTTCTCAACTTCTCGT	1 873
	1B-CDS-R1	GCTGAGACGAGTGCTAACC	1 873
	1B-CDS-F2	CATGTGGATGACAATGCTGAC	1 964
	1B-CDS-R2	CTTACTACTGATGCTGCCTGC	1 964
TaAGPL1-1D	1D-Pro-F	GGCCGGCGTCGCGAGAGCT	2 854
	1D-Pro-R	GAACATTTCTGTAAGTAACAC	2 854
	1D-CDS-F1	TACTGAGCTTTACAGAGGGCCG	2 040
	1D-CDS-R1	GACACAGTGAGGTAAGACCAG	2 040
	1D-CDS-F2	CATGTGGATGACAATGCTGAC	2 060
	1D-CDS-R2	CGGGAGTTTGTTTGCTCGCT	2 060
qTaAGPL1-1B	qTaAGPL1-F	GCCCCTGTTGGAGAGAGCCG	157
qTaAGPL1-1B	qTaAGPL1-R	TAGCAGGATCGTCTATGGCG	157
qActin	qActin-F	CCTCTCTTAGCACTTTCCAGCA	101
qActin	qActin-R	GTAAGTCCCCTTCACCGACTC	101
CAPS	CAPS-F	CATGTGCAGAAAGTGGACA	443
CAPS	CAPS-R	AGAAGGATCTGATTGTTCCA	443

品种 Variety	平均千粒重 Average thousand kernel weight/g	单倍型 Haplotype	品种 Variety	平均千粒重 Average thousand kernel weight/g	单倍型 Haplotype
烟农15号 Yannong 15	33.20	Hap1	新麦11号 Xinmai 11	43.27	Hap2
周麦20号 Zhoumai 20	46.20	Hap1	徐州24 Xuzhou 24	46.33	Hap2
濮麦9 Pumai 9	39.53	Hap1	洛麦21 Luomai 21	44.73	Hap2
陕229 Shan229	38.17	Hap1	丰德存麦1号Fengdecunmai 1	44.83	Hap2
泛麦8号 Fanmai 8	42.87	Hap1	周麦22号 Zhoumai 22	45.50	Hap2
豫麦49-198 Yumai 49-198	40.90	Hap1	周麦26号 Zhoumai 26	46.50	Hap2
豫麦2号 Yumai 2	37.20	Hap1	鲁麦23号 Lumai 23	46.37	Hap2
石家庄8号 Shijiazhuang 8	45.10	Hap1	驻麦4号 Zhumai 4	51.17	Hap2
百泉3039 Baiquan 3039	39.97	Hap1	众麦998 Zhongmai 998	49.30	Hap2
豫麦49 Yumai 49	37.00	Hap1	轮选66 Lunxuan 66	47.53	Hap2
藁2018 Gao 2018	37.97	Hap1	温9519 Wen 9519	47.17	Hap2
郑麦366 Zhengmai 366	36.80	Hap1	豫保1号 Yubao 1	46.47	Hap2
豫农908 Yunong 908	43.67	Hap1	许科1号 Xuke 1	46.33	Hap2
福穗3号 Fusui 3	42.07	Hap1	洛麦22 Luomai 22	47.60	Hap2
济南17号 Jinan 17	36.17	Hap1	平安8号 Ping’an 8	43.20	Hap2
石新828 Shixin 828	40.77	Hap1	郑麦9023 Zhengmai 9023	46.20	Hap2
稷麦336 Jimai 336	43.57	Hap1	泛麦5号 Fanmai 5	46.40	Hap2
洛麦47 Luomai 47	41.23	Hap1	许科316 Xuke 316	48.33	Hap2
鲁麦21号 Lumai 21	35.20	Hap1	徐麦856 Xumai 856	46.43	Hap2
科农199 Kenong 199	41.43	Hap1	郑农17 Zhengnong 17	46.83	Hap2
豫农982 Yunong 982	46.37	Hap1	中育12号 Zhongyu 12	45.00	Hap2
新麦26号 Xinmai 26	45.23	Hap1	烟辐188 Yanfu 188	50.53	Hap2

品种 Variety	平均千粒重 Average thousand kernel weight/g	单倍型 Haplotype	品种 Variety	平均千粒重 Average thousand kernel weight/g	单倍型 Haplotype
烟农15号 Yannong 15	33.20	Hap1	新麦11号 Xinmai 11	43.27	Hap2
周麦20号 Zhoumai 20	46.20	Hap1	徐州24 Xuzhou 24	46.33	Hap2
濮麦9 Pumai 9	39.53	Hap1	洛麦21 Luomai 21	44.73	Hap2
陕229 Shan229	38.17	Hap1	丰德存麦1号Fengdecunmai 1	44.83	Hap2
泛麦8号 Fanmai 8	42.87	Hap1	周麦22号 Zhoumai 22	45.50	Hap2
豫麦49-198 Yumai 49-198	40.90	Hap1	周麦26号 Zhoumai 26	46.50	Hap2
豫麦2号 Yumai 2	37.20	Hap1	鲁麦23号 Lumai 23	46.37	Hap2
石家庄8号 Shijiazhuang 8	45.10	Hap1	驻麦4号 Zhumai 4	51.17	Hap2
百泉3039 Baiquan 3039	39.97	Hap1	众麦998 Zhongmai 998	49.30	Hap2
豫麦49 Yumai 49	37.00	Hap1	轮选66 Lunxuan 66	47.53	Hap2
藁2018 Gao 2018	37.97	Hap1	温9519 Wen 9519	47.17	Hap2
郑麦366 Zhengmai 366	36.80	Hap1	豫保1号 Yubao 1	46.47	Hap2
豫农908 Yunong 908	43.67	Hap1	许科1号 Xuke 1	46.33	Hap2
福穗3号 Fusui 3	42.07	Hap1	洛麦22 Luomai 22	47.60	Hap2
济南17号 Jinan 17	36.17	Hap1	平安8号 Ping’an 8	43.20	Hap2
石新828 Shixin 828	40.77	Hap1	郑麦9023 Zhengmai 9023	46.20	Hap2
稷麦336 Jimai 336	43.57	Hap1	泛麦5号 Fanmai 5	46.40	Hap2
洛麦47 Luomai 47	41.23	Hap1	许科316 Xuke 316	48.33	Hap2
鲁麦21号 Lumai 21	35.20	Hap1	徐麦856 Xumai 856	46.43	Hap2
科农199 Kenong 199	41.43	Hap1	郑农17 Zhengnong 17	46.83	Hap2
豫农982 Yunong 982	46.37	Hap1	中育12号 Zhongyu 12	45.00	Hap2
新麦26号 Xinmai 26	45.23	Hap1	烟辐188 Yanfu 188	50.53	Hap2

品种 Variety	平均千粒重 Average thousand kernel weight/g	单倍型 Haplotype	品种 Variety	平均千粒重 Average thousand kernel weight/g	单倍型 Haplotype
石7221 Shi 7221	42.17	Hap1	潍麦8号 Weimai 8	47.83	Hap2
丰产3号 Fengchan 3	44.27	Hap1	徐麦30 Xumai 30	43.47	Hap2
舜麦1718 Shunmai 1718	43.07	Hap1	兰考矮早8 Lankaoaizao 8	49.13	Hap2
百农207 Bainong 207	42.47	Hap2	豫农416 Yunong 416	48.17	Hap2
富麦2008 Fumai 2008	48.13	Hap2	泰山21 Taishan 21	44.80	Hap2
赛德麦8号 Saidemai 8	46.27	Hap2	周麦16号 Zhoumai 16	45.37	Hap2
周麦36号 Zhoumai 36	45.27	Hap2	豫麦34号 Yumai 34	43.50	Hap2
西农20 Xinong 20	44.37	Hap2	泰山23号 Taishan 23	45.80	Hap2
中麦30 Zhongmai 30	46.00	Hap2	平安518 Ping’an 518	46.43	Hap2
百农4199 Bainong 4199	46.50	Hap2	光泰668 Guangtai 668	51.27	Hap2
郑麦139 Zhengmai 139	48.33	Hap2	新科麦170 Xinkemai 170	47.50	Hap2
淮麦18 Huaimai 18	42.20	Hap2	鑫华麦818 Xinhuamai 818	47.03	Hap2
周麦19号 Zhoumai 19	43.60	Hap2	中麦578 Zhongmai 578	48.50	Hap2
郑育麦9987 Zhengyumai 9987	50.90	Hap2	新麦60 Xinmai 60	46.93	Hap2
中麦875 Zhongmai 875	47.50	Hap2	国阳537 Guoyang 537	46.53	Hap2
汝州0319 Ruzhou 0319	47.40	Hap2	航宇19 Hangyu 19	43.53	Hap2
许农5号 Xunong 5	46.23	Hap2	新麦45 Xinmai 45	43.53	Hap2
许科718 Xuke 718	48.57	Hap2	周麦44 Zhoumai 44	46.40	Hap2
商麦178 Shangmai 178	45.20	Hap2	洲元9369 Zhouyuan 9369	36.47	Hap2
科兴3302 Kexing 3302	48.33	Hap2	平安6号 Ping’an 6	41.40	Hap2
鲁原502 Luyuan 502	43.47	Hap2	百3217 Bai 3217	37.30	Hap2
平麦998 Pingmai 998	43.30	Hap2	连麦2号 Lianmai 2	39.27	Hap2