Analysis of Genetypes Related to Important Agronomic Traits of Purple Waxy Special Rice Variety Ziliangyou 737
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摘要:
目的 解析紫糯特种稻紫两优737及其亲本的产量、品质相关性状的基因型构成，为该品种及其双亲在育种和生产上的进一步应用提供分子依据。 方法 应用10个重要基因的13个分子标记，分析紫两优737及其亲本紫392S、福恢737在产量、品质等相关基因座位上的等位基因变化。 结果 在7个基因座位上，紫两优737及其亲本均携带相同等位基因型。在穗实粒数主效基因Gn1a座位上均携带高产等位基因Ha-Gn1a；在粒重和粒长主效基因GS3座位上均携带长粒型等位基因MH-GS3；在种皮基因Pb座位上均携带紫色果皮等位基因型；在糯稻蜡质基因wx座位上均携带低直链淀粉含量等位基因型；此外在淀粉分支酶基因SBE1和SBE3以及极限糊精酶基因PUL基因座位上亦均携带相同等位基因型。在3个基因座位上，双亲携带不同等位基因型。在Hd3a座位上，紫392S、福恢737和紫两优737分别携带Hd3aNip、hd3aKasa和Hd3aNip/hd3aKasa；在Hd1座位上，分别携带Hd1jap、Hd1ind和Hd1jap/Hd1ind；在焦磷酸化酶大亚基基因AGPlar座位上，分别为Ⅰ型，Ⅱ型及Ⅰ/Ⅱ杂合型。 结论 紫两优737重要农艺性状的遗传解析为其具有高产、优质、适应性好及种皮颜色等特点提供了分子证据，并为相关有利等位基因的进一步利用提供参考。 Abstract:Objective This study was conducted to analyze the genotypes of yield and quality-related traits of the purple waxy special rice Ziliangyou 737 and its parents, and to provide molecular basis for the further application of this variety and its parents in breeding and production. Method Using 13 molecular markers of 10 important genes, the alleles of Ziliangyou 737 and its parents Ziliang392s and Fuhui 737 were analyzed in relation to yield and quality. Result In 7 gene loci, both Zizliangyou 737 and its parents carry the same allele genotype. The high-yield allele Ha-Gn1a was carried in the seat of Gn1a, the major gene of panicles number. The long grain allele MH-GS3 was carried in the GS3 locus of the major gene for grain weight and grain length. The alleles of purple pericarp were carried in the Pb locus of seed coat gene. All alleles with low amylose content in waxy gene wx locus were carried in glutinous rice. In addition, the same allelic genotype was carried in the starch branch enzyme genes SBE1 and SBE3 and the limit dextrin enzyme gene PUL gene locus. In three loci, parents carry different alleles. In the Hd3a seat, the Zi 392S, Fuhui 737 and Ziliangyou 737 carry Hd3aNip, hd3aKasa and Hd3aNip/hd3aKasa respectively. On the Hd1 seat, carry Hd1jap, Hd1ind and Hd1jap/Hd1ind respectively. In the large subunit gene AGPlar locus of pyrophosphorylase, there were type I, type II and hybrid type I/II, respectively. Conclusion The genetic analysis of important agronomic traits of Ziliangyou 737 provided molecular evidence for its high yield, high quality, good adaptability and seed coat color, and provided reference for further utilization of related favorable alleles -
Key words:
- Purple waxy hybrid rice /
- molecular markers /
- genotyping /
- high yield /
- high quality
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图 1 水稻产量因子Gn1a、 GS3基因相关标记对紫两优737及其亲本的检测结果
A：Gn1a-M1。M：20bp DNA Marker；1：明恢63；2：宜优673；3：紫392S；4：福恢737；5：紫两优737。B： GS3-PstⅠ，原始扩增产物（左）和酶切产物（右）。M：100 bp DNA Marker； 1：珍汕97，2：明恢63，3：紫392S，4：福恢737；5：紫两优737。
Figure 1. Detection of Ziliangyou 737 and its parents with markers for rice yield components Gn1a and GS3
A: Gn1a-M1. M: 20bp DNA Marker; 1: Minghui 63; 2: Yiyou 673; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737. B: GS3-PstⅠ, original amplified products(left) and enzyme digestion products (right). M: 100 bp DNA Marker; 1: Zhengshan 97; 2: Minghui 63; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737.
图 2 水稻Hd3a、 Hd1基因相关标记对紫两优737及其亲本的检测结果
A： hd3afnp。M：100 bp DNA Marker；1：日本晴；2：Kasalath；3：紫392S；4：福恢737；5：紫两优737。B：Si9337。M：20 bp DNA Marker；1：日本晴；2：9311；3：紫392S；4：福恢737；5：紫两优737。
Figure 2. Detection of Ziliangyou 737 and its parents with markers for Hd3a and Hd1
A: hd3afnp. M: 100 bp DNA Marker; 1: Nipponbare; 2: Kasalath; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737. B: Si9337. M: 20 bp DNA Marker; 1: Nipponbare; 2: 9311; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737.
图 3 糯稻蜡质基因相关标记对紫两优737及其亲本的检测结果
A：We 2-2。M：20 bp DNA Marker；1：荆糯6；2：9311；3：紫392S；4：福恢737；5：紫两优737。B：Wx-t1。M：20 bp DNA Marker；1：荆糯6；2：9311；3：紫392S；4：福恢737；5：紫两优737。C：484/W2R，原始扩增产物（左）和酶切产物（右）；M：100 bp DNA Marker；1：9311；2：明恢63；3：紫392S；4：福恢737；5：紫两优737。D：PCR-AccⅠ，原始扩增产物（左）和酶切产物（右）；M：100 bp DNA Marker；1：9311；2：明恢63；3：紫392S；4：福恢737；5：紫两优737。
Figure 3. Detection of Ziliangyou 737 and its parents with markers for wx
A: We 2-2. M: 20 bp DNA Marker; 1: Jingnuo 6; 2: 9311; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737. B: Wx-t1. M: 20 bp DNA Marker; 1: Jingnuo 6; 2: 9311; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737. C: 484/W2R, original amplified products (left) and enzyme digestion products (right); M: 100 bp DNA Marker; 1: 9311; 2: Minghui 63; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737. D: PCR-AccⅠ, original amplified products (left) and enzyme digestion products (right); M: 100 bp DNA Marker; 1: 9311; 2: Minghui 63; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737.
图 4 水稻淀粉合成相关基因、紫色种皮基因Pb相关标记对紫两优737及其亲本的检测结果
A：Sbe1。M：100 bp DNA Marker。1：9311；2：明恢63；3：紫392S；4：福恢737；5：紫两优737。B：AGPlar M1。M：20 bp DNA Marker；1：珍汕97B；2：9311；3：紫392S；4：福恢737；5：紫两优737。C：PUL M2。M：20 bp DNA Marker；1：9311；2：宜优673；3：紫392S；4：福恢737；5：紫两优737。D：RS/SpeⅠ，原始扩增产物（左）和酶切产物（右）。M：100 bp DNA Marker；1：密阳23；2：宜优673；3：紫392S；4：福恢737；5：紫两优737。E：CAPSPb，原始扩增产物（左）和酶切产物（右）。M：100 bp DNA Marker；1：日本晴； 2：9311；3：紫392S；4：福恢737；5：紫两优737。
Figure 4. Detection of Ziliangyou 737 and its parents with markers for starch synthesis-related genes and Pb gene
A: Sbe1. M: 100 bp DNA Marker. 1: 9311; 2: Minghui 63; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737. B: AGPlar M1. M: 20 bp DNA Marker; 1: Zhenshan 97B; 2: 9311; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737. C: PUL M2. M: 20 bp DNA Marker; 1: 9311; 2: Yiyou 673; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737. D: RS/SpeⅠ, original amplified products (left) and enzyme digestion products (right). M: 100 bp DNA Marker; 1: Miyang 23; 2: Yiyou 673; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737. E: CAPSPb, original amplified products (left) and enzyme digestion products (right). M: 100 bp DNA Marker; 1: Nipponbare; 2: 9311; 3: Zi 392S; 4: Fuhui 737; 5: Ziliangyou 737.
表 1 检测10个基因的相关标记信息
Table 1. Markers’ information for 10 genes
基因
Gene标记
Marker引物序列
Primer sequence (5′-3′)退火温度
Annealing temperature/℃等位基因参照
Allele referenceGn1a Gn1a-M1[21] CTCTTGCTTCATTATCAATC 55 明恢63 Minghui 63 AAACTACACAAGAATCTGCT GS3 GS3-PstⅠ[21]
（限制性内切酶 Restriction enzyme：PstⅠ）TATTTATTGGCTTGATTTCCTGTG 55 珍汕97 Zhenshan 97，明恢63 Minghui 63 GCTGGTTTTTTACTTTCATTTGCC Hd3a hd3afnp inner[22] AGCGGCAGGAGaGTCTACAA 62 日本晴 Nipponbare， Kasalath TCaGGATCATCGTTAGCTAGGG hd3afnp outer AAtCGAGGGGAGTATATTGCTAGT GCTaCATGAGAGACCTTAGCCTT Hd1 Si9337[23] AGATGTCCCTTCACTTCAGC 60 9311，日本晴 Nipponbare CGAAACGGCCCTTGATCC wx We 2-2[24] CACTACAAGACACACTTGCAC 55 荆糯6 Jingnuo 6, 9311 GTCATCTAGCCCACCACCTT Wx-t1[25] ATGTCGGCTCTCACCACG 55 荆糯6 Jingnuo 6, 9311 ACCGACCGCTGCTGCTTG 484/W2R[26]
（限制性内切酶 Restriction enzyme：AccⅠ）CTTTGTCTATCTCAAGACAC 55 9311，明恢63 Minghui 63 TTTCCAGCCCAACACCTTAC PCR- AccⅠ [27]
（限制性内切酶 Restriction enzyme：AccⅠ）GCTTCACTTCTCTGCTTGTG 55 日本晴Nipponbare, 明恢63 Minghui 63 ATGATTTAACGAGAGTTGAA Sbe1 Sbe1[28] GAGTTGAGTTGCGTCAGATC 57 9311，日本晴 Nipponbare AATGAGGTTGCTTGCTGCTG sbe3-rs RS/SpeⅠ[29]
（限制性内切酶 Restriction enzyme：SpeⅠ）ATGTGATGTGCTGGATTTGG 55 密阳 23 Miyang 23，宜优673 Yiyou 673 TGTGGTTTTCATACCGTTCTTA AGPlar AGPlar M1[30] CGTTCAGGTTCAGGCAATCA 58 珍汕97 Zhenshan 97, 9311 GGAAGGGTGGTGATGTGGAG PUL PUL M2[30] GACAACCGTCCGCTTTAGTTTC 58 9311, 宜优673 Yiyou 673 GCATTTGAGAGGGTTTGGATTC Pb CAPSPb [31]
（限制性内切酶 Restriction enzyme：BamHⅠ）AAATCAGTTGTCCCGTCCA 58 9311，日本晴 Nipponbare TTAGGGAGTTGGTGATGGG 
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表 2 紫两优737在云南、福建和安徽区域试验产量表现[1,20]
Table 2. Yield performance of Ziliangyou 737 in regional trails in Yunnan, Fujian and Anhui
试验类型
Type of test品种
Varieties平均产量
Average Yield/（kg·hm−2）比对照增减产
Ratio compared with CK/%云南区试 Reginal trial in Yunnan 紫两优737 Ziliangyou 737 7566.90 152.1.0 癸能紫米 Guinengzimi（CK） 3002.10 福建区试 Reginal trial in Fujian 紫两优737 Ziliangyou 737 7505.48 −3.82 宜优673 Yiyou 673（CK） 7804.07 安徽区试 Reginal trial in Anhui 紫两优737 Ziliangyou 737 8441.63 −0.16 Ⅱ优838（CK） 8456.55
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表 3 紫两优737在云南、福建区域试验稻米品质[1]
Table 3. Rice quality of Ziliangyou 737 in regional trails in Yunnan and Fujian
类别 Types 糙米率
Brown rice percentage/%精米率
Head rice percentage/%整精米率
Head rice percentage/%粒长
Grain length/mm长宽比 Ratio of grain length to width 直链淀粉 Amylase content/% 碱消值 Alkali value 胶稠度
Gel consistency/mm云南区试Regional trial in Yunnan 78.4 69.4 55.3 6.3 2.7 2.6 7.0 90 福建区试 Reginal trial of in Fujian 79.3 68.5 65.1 6.4 2.9 2.1 7.0 97
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