柳　娜，曹　東，王世紅，楊文雄

(1.甘肅省農(nóng)業(yè)科學(xué)院 小麥研究所，蘭州　730070；2.甘肅農(nóng)業(yè)大學(xué) 農(nóng)學(xué)院，蘭州　730070)

?

104份甘肅小麥品種主要品質(zhì)基因的分子標(biāo)記檢測(cè)

柳娜1，曹東2，王世紅1，楊文雄1

(1.甘肅省農(nóng)業(yè)科學(xué)院 小麥研究所，蘭州730070；2.甘肅農(nóng)業(yè)大學(xué) 農(nóng)學(xué)院，蘭州730070)

摘要高分子量麥谷蛋白亞基、黃色素質(zhì)量分?jǐn)?shù)和1B/1R易位系均影響小麥的加工品質(zhì)。為明確相關(guān)品質(zhì)性狀基因在104份甘肅育成小麥品種中的分布，并為小麥品質(zhì)育種提供親本材料，利用高分子質(zhì)量谷蛋白亞基 Dx5、 Bx7、 By8、 By9和黃色素質(zhì)量分?jǐn)?shù)基因及1B/1R易位系的特異性標(biāo)記對(duì)104份小麥品種進(jìn)行檢測(cè)。結(jié)果表明，含基因 Dx5、 Bx7、 By8、 By9和1B/1R易位系的品種分別占總品種數(shù)的63.46%、76.92%、52.88%、45.19%和45.19%。含有等位基因 Psy-A1a和 Psy-A1b品種分別占總品種數(shù)的60.58%和21.15%，其黃色素質(zhì)量分?jǐn)?shù)均值差異達(dá)到顯著水平(P<0.05)?？傮w來看，甘肅育成小麥品種中含適合加工饅頭和面粉等傳統(tǒng)食品的單個(gè)品質(zhì)優(yōu)質(zhì)基因頻率較高，而優(yōu)質(zhì)基因組合的比例較低，具有黃色素質(zhì)量分?jǐn)?shù)較高的品種比例高。

關(guān)鍵詞甘肅小麥；高分子量谷蛋白亞基；品質(zhì)基因；分子標(biāo)記

小麥面筋強(qiáng)度及其面制品色澤等主要受高分子量谷蛋白亞基(HMW-GS)、1B/1R易位系、黃色素質(zhì)量分?jǐn)?shù)等因素的影響。已有研究表明，HMW-GS是影響小麥面團(tuán)特性和面筋強(qiáng)度的關(guān)鍵因素，而且其不同類別的亞基對(duì)小麥品質(zhì)改良的貢獻(xiàn)各異[1-2]。Payne等[3]和Branlard等[4]研究表明， Glu-D1位點(diǎn)上小麥烘烤等品質(zhì)表現(xiàn)最優(yōu)的亞基為Dx5+Dy10，并可利用 Dx5標(biāo)記準(zhǔn)確快速的檢測(cè)和判斷小麥品種中是否含有Dx5+Dy10優(yōu)質(zhì)亞基。D’Ovideo等[5-6]和Bustos等[7]設(shè)計(jì)了一系列可以區(qū)分基因 1Dx5、 1Dx2、 1Dx3、 1Dx4、 1Dy10、 1Dy12和Null等的HMW-GS的特異引物。Ma等[8]和張曉科等[9]分別建立了基因 Ax2、 Bx17和 Dx5的多重PCR反應(yīng)體系。Bx7亞基在小麥中分布較廣，通常與亞基By8或By9連鎖，但亞基組合Bx7+By8對(duì)面筋強(qiáng)度的貢獻(xiàn)大于其Bx7+By9[5]。1B/1R易位系即小麥的1B染色體短臂被黑麥的1R染色體短臂取代而形成[10]。在中國(guó)小麥育種進(jìn)程中1B/1R易位系得到廣泛應(yīng)用，同時(shí)也推進(jìn)小麥育種進(jìn)程[11]。Francis等[12]已開發(fā)檢測(cè)1B/1R易位系的基因標(biāo)記，可準(zhǔn)確快速方便的鑒定其是否攜帶1B/1R易位系。但與此同時(shí)也產(chǎn)生一些問題，如1B/1R易位品種的面筋質(zhì)量下降，使小麥的加工品質(zhì)變劣[10,13-15]。因此，對(duì)于品質(zhì)育種而言，判斷其親本及后代中是否含有1B/1R易位是至關(guān)重要的。中國(guó)傳統(tǒng)面條和饅頭等面制品需亮度好和低黃色素質(zhì)量分?jǐn)?shù)的小麥面粉來加工。因此，研究和選育低黃色素質(zhì)量分?jǐn)?shù)的新品種是中國(guó)小麥品質(zhì)育種的主要目標(biāo)。而小麥籽粒中黃色素質(zhì)量分?jǐn)?shù)是多基因調(diào)控的，研究表明，染色體7A和7B上的基因?qū)ζ湄暙I(xiàn)較大[16-17]。He等[18]開發(fā)了位于7A染色體的黃色素基因標(biāo)記 YP7A，能有效區(qū)分小麥7A染色體控制高、低黃色素質(zhì)量分?jǐn)?shù)的等位基因 Psy-A1a和 Psy-A1b。Parker等[19]研究得出位于小麥染色體3A和7A上控制黃色素質(zhì)量分?jǐn)?shù)的主效QTL可分別解釋其表型變異的13%和60%。以上相關(guān)品質(zhì)基因分子標(biāo)記的開發(fā)為小麥品質(zhì)改良提供可靠的理論依據(jù)。

高鳳梅等[20]、蘆靜等[21]、王憲國(guó)等[22]和胡鳳靈等[23]分別對(duì)黑龍江、新疆、寧夏及全國(guó)冬、春小麥的相關(guān)品質(zhì)基因進(jìn)行檢測(cè)，掌握其相關(guān)品質(zhì)優(yōu)質(zhì)基因的分布情況。但有關(guān)甘肅省小麥品質(zhì)相關(guān)基因檢測(cè)的研究較缺乏。本研究選用甘肅省育成104份小麥品種資源，利用 Dx5、 YP7A、 Bx7、 By8 、 By9和 1B/1R特異性標(biāo)記進(jìn)行分子檢測(cè)，掌握甘肅育成品種中所含相關(guān)品質(zhì)基因的種類及分布情況，為甘肅省小麥品質(zhì)育種提供有價(jià)值的親本材料，加快品質(zhì)改良進(jìn)程。

1材料與方法

1.1材 料

供試品種為104份甘肅省育成審定的小麥品種，由甘肅省農(nóng)業(yè)科學(xué)院小麥課題組保存。并在2012-2013年種植于甘肅省武威市黃羊鎮(zhèn)甘肅農(nóng)科院小麥育種基地，常規(guī)管理，成熟時(shí)收獲。這些冬小麥(72份)和春小麥(32份)基本反映甘肅省近10 a小麥育種現(xiàn)狀。

1.2試驗(yàn)方法

1.2.1DNA提取選取每個(gè)品種新鮮幼嫩的小麥黃化苗，用CTAB法[24-25]提取小麥基因組DNA。為保證檢測(cè)結(jié)果的準(zhǔn)確性，每個(gè)品種提取3批DNA，并分別檢測(cè)。

1.2.2PCR擴(kuò)增與電泳檢測(cè)以D’Ovidio等[5]、He等[18]、Ma等[26]、Lei等[27]和Francis等[12]開發(fā)的功能標(biāo)記 Dx5、 YP7A、 Bx7、 By9、 By8和 1B/1R進(jìn)行分子檢測(cè)。引物由上海生工生物工程技術(shù)服務(wù)有限公司合成(表1)。擴(kuò)增體系10 μL，含Tris-HCl 20 mmol/L(pH 8.4)，KCl 20 mmol/L，dNTPs 200 μmol/L，MgCl21.5 mmol/L，引物10 pmol/L，1.5 UTaqDNA 聚合酶，模板DNA為50 ng。

最后擴(kuò)增產(chǎn)物用15 g/L的瓊脂糖凝膠電泳檢測(cè)，緩沖液為 1×TAE 溶液，160 V電壓電泳45 min，染色、掃描成像。

表1　引物序列、擴(kuò)增片段及對(duì)應(yīng)的基因型

1.2.3黃色素質(zhì)量分?jǐn)?shù)測(cè)定采用已改進(jìn)AACC14-50 法[28]，取各個(gè)小麥品種的面粉樣品3 g，加入提取液水飽和正丁醇15 mL，于振蕩器上振蕩浸提1 h后，靜置和離心。用分光光度計(jì)(波長(zhǎng)為440 nm)測(cè)定吸光度，重復(fù)3次，取其平均值。

2結(jié)果與分析

2.1甘肅小麥中部分HMW-GS的分布

利用部分HWM-GS特異性引物檢測(cè)可以得出，在 Glu-D1位點(diǎn)，標(biāo)記 Dx5在含亞基Dx5的品種中可擴(kuò)增出450 bp的條帶(圖1)。在 Glu-B1位點(diǎn)，標(biāo)記 Bx7在含亞基Bx7的品種中可擴(kuò)增出630 bp和766 bp的2條帶型(圖2)；標(biāo)記 By8在含亞基By8的品種中可擴(kuò)增出527 bp的條帶(圖3)；標(biāo)記 By9在含亞基By9的品種中可擴(kuò)增出662 bp的帶型(圖4)。

在104份甘肅省育成的小麥品種中，含亞基 Dx5、 Bx7、 By8和 By9的品種分別有66、80、55和88份，分別占總品種數(shù)的63.46%、76.92%、52.88%和45.19%。其中，所有冬小麥品種中含亞基Dx5、Bx7、By8和By9分別占總品種數(shù)的63.89%、77.78%、55.56%和41.67%；而在所有春小麥品種中的頻率依次為62.5%、75%、46.88%和53.13%(表2)。

M.DNA ladder 2000；1.隴春23號(hào)Longchun 23；2.隴春28號(hào)Longchun 28；3.隴春22號(hào)Longchun 22；4.武春5號(hào)Wuchun 5；5.武春6號(hào)Wuchun 6；6.武春7號(hào)Wuchun 7；7.武春8號(hào)Wuchun 8；8.甘育1號(hào)Ganyu 1；9.平?jīng)?3號(hào)Pingliang 43；10.平?jīng)?4號(hào)Pingliang 44；11.涇麥1號(hào)Jingmai 1；12.天選43Tianxuan 43；13.隴春26號(hào)Longchun 26

圖1 Dx5標(biāo)記擴(kuò)增部分品種瓊脂糖凝膠電泳

Fig.1Agarose gel electrophoresis of some tested materials amplified by Dx5

M.DNA ladder 2000；1.隴春23號(hào)Longchun 23；2.隴春28號(hào)Longchun 28；3.隴春30號(hào)Longchun 30；4.臨農(nóng)826Linnong 826；5.隴鑒103Longjian 103；6.環(huán)冬3號(hào)Huandong 3；7.隴育1號(hào)Longyu 1；8.武春8號(hào)Wuchun 8；9.臨麥33Linmai 33；10.天選50Tianxuan 50；11.隴鑒9343Longjian 9343；12.張冬30號(hào)Zhangdong 30；13.靜麥10號(hào)Jingmai 10

圖2 Bx7標(biāo)記擴(kuò)增部分品種瓊脂糖凝膠電泳

Fig.2Agarose gel electrophoresis of some tested materials amplified by Bx7

M.DNA ladder 2000；1.隴鑒9811Longjian 9811；2.武春3號(hào)Wuchun 3；3.張冬30號(hào)Zhangdong 30；4.隴鑒301Longjian 301；5.銀春8號(hào)Yinchun 8；6.靈臺(tái)3號(hào)Lingtai 3；7.西峰28號(hào)Xifeng 28；8.定豐16號(hào)Dingfeng 16；9.中梁23號(hào)Zhongliang 23；10.張春21號(hào)Zhangchun 21；11.隴育2號(hào)Longyu 2；12.隴輻2號(hào)Longfu 2

圖3 By8標(biāo)記擴(kuò)增部分品種瓊脂糖凝膠電泳

Fig.3 Agarose gel electrophoresis of some tested materials amplified by By8

2.2甘肅小麥黃色素質(zhì)量分?jǐn)?shù)等位基因的檢測(cè)

2.2.1 Psy-A1位點(diǎn)變異檢測(cè)利用標(biāo)記 YP7A對(duì)104份甘肅小麥品種中 Psy-A1位點(diǎn)的等位變異檢測(cè)，在含 Psy-A1a和 Psy-A1b基因的品種中可分別擴(kuò)增出194 bp和231 bp的2條帶型(圖5)。結(jié)果顯示，104份甘肅小麥中有63份擴(kuò)增出194 bp片段，占總品種數(shù)的60.58%，含 Psy-A1a基因；22份品種中擴(kuò)增出231 bp片段，占總品種數(shù)的21.15%，含 Psy-A1b基因，與低黃色素質(zhì)量分?jǐn)?shù)相關(guān)[29]。在冬小麥品種中，44份(61.11%)品種中含有 Psy-A1a基因，17份(23.61%)品種含有 Psy-A1b基因；在春小麥品種中，19份(59.38%)品種中含有 Psy-A1a基因，5份(15.63%)品種含有 Psy-A1b基因(表3)。

M.DNA ladder 2000；1.武春3號(hào)Wuchun 3；2.武春6號(hào)Wuchun 6；3.武春7號(hào)Wuchun 7；4.定豐12號(hào)Dingfeng 12；5.甘育1號(hào)Ganyu 1；6.定豐10號(hào)Dingfeng 10；7.中天1號(hào)Zhongtian 1；8.隴育4號(hào)Longyu 4；9.臨麥32號(hào)Linmai 32；10.武都17號(hào)Wudu 17；11.靜麥2號(hào)Jingmai 2；12.航選01Hangxuan 01

圖4　 By9標(biāo)記擴(kuò)增部分供試材料瓊脂糖凝膠電泳

M.DNA ladder 2000；2.武春3號(hào)Wuchun 3；3.武春7號(hào)Wuchun 7；4.武春5號(hào)Wuchun 5；5.靈臺(tái)2號(hào)Lingtai 2；6.環(huán)冬4號(hào)Huandong 4；7.隴育2號(hào)Longyu 2；8.西峰28Xifeng 28；9.環(huán)冬3號(hào)Huandong 3；10.中梁23Zhongliang 23；11.中梁24Zhongliang 24；12.天選45Tianxuan 45

圖5　 YP7A標(biāo)記擴(kuò)增部分品種瓊脂糖凝膠電泳

2.2.2 Psy-A1基因與黃色素質(zhì)量分?jǐn)?shù)的關(guān)系從表4可以看出，大多含有 Psy-A1a 基因的小麥品種黃色素質(zhì)量分?jǐn)?shù)較高，均值為3.5 mg/kg，而含有 Psy-A1b基因的品種所對(duì)應(yīng)的黃色素質(zhì)量分?jǐn)?shù)較低，均值為2.1 mg/kg。依據(jù)t測(cè)驗(yàn)結(jié)果可以看出，分別含 Psy-A1a和 Psy-A1b 2種基因的品種間黃色素質(zhì)量分?jǐn)?shù)均值差異顯著。

2.3甘肅小麥1B/1R易位系的分布

利用1B/1R易位系的特異性標(biāo)記對(duì)104份甘肅小麥品種進(jìn)行檢測(cè)，可擴(kuò)增出1 500 bp的條帶(圖6)。結(jié)果顯示，在104份小麥品種中有47份材料含有1B/1R易位系，占總品種數(shù)的45.19%。其中，冬小麥和春小麥分別有32份和15份，頻率分別為44.44%和46.88%。

含有Dx5亞基的小麥品種面筋強(qiáng)度高；同時(shí)具有低黃色素質(zhì)量分?jǐn)?shù)的品種有利于面條和饅頭等面制品色澤的改善。在甘肅省育成的104份冬、春小麥中，有‘隴春22號(hào)’、‘靜麥10號(hào)’、‘隴鑒9343’、‘蘭天17’等37份(35.58%)小麥含有優(yōu)質(zhì)亞基Dx5，且不是1B/1R易位系。同時(shí)含亞基Dx5和 Psy-A1b基因，且不是1B/1R易位系的有‘隴春22號(hào)’和‘定西38號(hào)’等11份(10.58%)小麥品種。由此可以得出，具有多種優(yōu)質(zhì)基因的小麥品種較少。

表4　104份甘肅小麥品種黃色素質(zhì)量分?jǐn)?shù)t測(cè)驗(yàn)

注：不同字母表示差異達(dá)顯著水平(P<0.05)。

Note:Different letters indicate significant differences (P<0.05).

M.DNA ladder 2000；2.武春3號(hào)Wuchun 3；3.武春7號(hào)Wuchun 7；4.武春5號(hào)Wuchun 5；5.靈臺(tái)2號(hào)Lingtai 2；6.環(huán)冬4號(hào)Huandong 4；7.隴育2號(hào)Longyu 2；8.西峰28Xifeng 28；9.環(huán)冬3號(hào)Huandong 3；10.中梁23Zhongliang 23；11.中梁24Zhongliang 24；12.天選45Tianxuan 45

圖61B/1R標(biāo)記擴(kuò)增部分供試材料瓊脂糖凝膠電泳

Fig.6Agarose gel electrophoresis of some tested materials amplified by 1B/1R

3討 論

近年來品質(zhì)育種越來越受到許多小麥育種家的重視。不同面制品對(duì)其品質(zhì)的要求各異，傳統(tǒng)的面食品如面條和饅頭等要求色白[30]。研究表明，部分HMW-GS和LMW-GS對(duì)改良小麥面筋強(qiáng)度具有重要的作用，特別是優(yōu)質(zhì)亞基Dx5+Dy10等的引入，改善了面筋質(zhì)量[31]。本研究用功能性標(biāo)記 Dx5、 YP7A、 Bx7、 By9、 By8和 1B/1R 易位系的特異性標(biāo)記對(duì)甘肅省近年來審定的104份小麥品種進(jìn)行檢測(cè)，含Dx5、Bx7、By8和By9亞基的頻率依次為63.46%、76.92%、52.88%和45.19%。與其他地區(qū)相比[20,32-33]，含亞基Dx5、By8、 By9和1B/1R易位系的分布頻率高于其他麥區(qū)，而含Bx7亞基的低于其他麥區(qū)。由此可以看出，含單個(gè)品質(zhì)優(yōu)質(zhì)亞基的分布頻率較高，這些基因在甘肅冬春麥中的分布不同，冬小麥所含的頻率高于春小麥。但為今后更好地改善小麥品種的面筋強(qiáng)度，更應(yīng)繼續(xù)引進(jìn)含優(yōu)質(zhì)基因品種，并結(jié)合相關(guān)品質(zhì)基因標(biāo)記輔助選擇。在生產(chǎn)中，由于大多數(shù)冬小麥品種的品質(zhì)優(yōu)于春小麥品種，冬小麥品質(zhì)改良重視程度高于春小麥，在小麥生產(chǎn)和加工中傾向于使用冬小麥品種。

小麥品種中黃色素質(zhì)量分?jǐn)?shù)的高低與面粉及面制品的色澤緊密相關(guān)[32]。甘肅省育成小麥品種表現(xiàn)出 Psy-A1a頻率(60.58%)偏高，需加強(qiáng)對(duì)小麥色澤的選擇，提高面粉白度來滿足人們的需求。小麥品質(zhì)育種應(yīng)不宜采用含1B/1R易位系的品種作親本培育優(yōu)質(zhì)新品種[34]。而在104份甘肅小麥中，1B/1R易位系品種(45.19%)所占的比例較高。

綜上可以得出，在甘肅育成品種中含Dx5、Bx7、By8和By9亞基的頻率較高；同時(shí)含優(yōu)質(zhì)亞基Dx5和 Psy-A1b基因的小麥品種較少。說明近年來甘肅小麥品質(zhì)育種比較單一，今后應(yīng)緊密結(jié)合分子標(biāo)記輔助選擇，選擇攜帶優(yōu)質(zhì)基因的品種充當(dāng)親本，聚合多個(gè)品質(zhì)優(yōu)質(zhì)基因，加快品質(zhì)育種進(jìn)程，改善小麥品質(zhì)。

參考文獻(xiàn)Reference：

[1]PAYNE P I.Genetics of wheat storage protein and the effect of allelic variation on bread-making quality[J].AnnualReviewofPlantPhysiology,1987,38:141-153.

[2]LIU L,HE Z H,YAN J,etal.Allelic variation at the Glu1- and Glu3-loci,presence of the 1B·1R translocation,and their effects on mixographic properties in Chinese bread wheats[J].Euphytica,2005,142:197-204.

[3]PAYNE P I,NIGHTINGALE M A,KRATTIGER A F,etal.The relationship between HMW glutenin subunits composition and the bread-making quality of British grown wheat varieties[J].JournalofAgriculturalandFoodChemistry,1987,40:51-65.

[4]BRANLARD G,DARDEVET R,SACCOMANO F,etal.Genetics diversity of wheat storage proteins and bread wheat quality[J].Euphytica,2001,119:59-67.

[5]D’OVIDIO R,ANDERSON O D.PCR analysis to distinguish between alleles of a member of a multigene family correlated with wheat bread-making quality[J].TheoreticalandAppliedGenetics,1994,88(6-7):759-763.

[6]D’OVIDIO R,MASCI S,PORCEDU D.Development of a set of oligonucleotide primers specific for genes at the Glu-1 complex loci of wheat[J].TheoreticalandAppliedGenetics,1995,91(2):189-194.

[7]BUSTOS A DE.Molecular characterization of the inactive allele of the gene Glu-A1 and the development of a set of AS-PCR makers of HMW glutenins of wheat[J].TheoreticalandAppliedGenetics,2000,100:1085-1094.

[8]MA W,DAGGARD G,SUTHERLAND M.Molecular Marker for Quality Attributes in Wheat [C]//Proceedings of the Ninth Assembly.Australia:Wheat Breeding Society,1999:115-117.

[9]張曉科,夏先春,王忠偉,等.小麥品質(zhì)性狀分子標(biāo)記多重PCR體系的建立[J].作物學(xué)報(bào), 2007,33(10):1703-1710.

ZHANG X K,XIA X CH,WANG ZH W,etal.Establishment of multiplex-PCR for quality traits in common wheat [J].ActaAgronomicaSinica,2007,33(10):1703-1710(in Chinese with English abstract).

[10]劉建軍,何中虎,PENA R J,等.1BL/1RS 易位對(duì)小麥加工品質(zhì)的影響[J].作物學(xué)報(bào),2004,30(2):149-153.

LIU J J,HE ZH H,PENA R J,etal.Effect of 1BLP1RS translocation on grain quality and noodle quality in bread wheat[J].ActaAgronomicaSinica,2004,30(2):149-153(in Chinese with English abstract).

[11]周陽(yáng),何中虎,張改生,等.1BL/1RS 易位系在我國(guó)小麥育種中的應(yīng)用[J].作物學(xué)報(bào),2004,30(6):531-535.

ZHOU Y,HE ZH H,ZHANG G SH,etal.Utilization of 1BL/1RS translocation in wheat breeding in China [J].ActaAgronomicaSinica,2004,30(6):531-535(in Chinese with English abstract).

[12]FRANCIS H A,LEITCH A R,KOEBNER R M D.Conversion of a RAPD-generated PCR product containing a novel dispersed repetitive element into a fast and robust assay for the presence of rye chromatin in wheat[J].TheoreticalandAppliedGenetics,1995,90(5):636-642.

[13]GRAYBOSCH R A,PETERSON C J,HANSEN L E,etal.Relationships between protein solubility characteristics,1BL/1RS,high-molecular weight glutenin composition and end-use quality in winter wheat germplasm[J].CerealChemistry,1990,67:342-349.

[14]DHALIWAL A S,MARES D J,MARSHALL D R.Effect of 1B/1R chromosome translocation on milling and quality characteristics of bread wheats[J].CerealChemistry,1987,64:72-76.

[15]MARTIN P,GOMEZ M,CARRILLO J M.Interaction between allelic variation at the Glu-D1 locus and a 1BL/1RS translocation on flour quality in bread wheat[J].CropScience,2001,41:1080-1084.

[16]張立平,閻俊,夏先春,等.普通小麥籽粒黃色素含量的QTL分析[J].作物學(xué)報(bào),2006,32(1):41-45.

ZHANG L P,YAN J,XIA X CH,etal.QTL mapping for kernel yellow pigment content in common wheat[J].ActaAgronomicaSinica,2006,32(1):41-45(in Chinese with English abstract).

[17]KUCHEL H,LANGRIDGE P,MOSIONEK L,etal.The genetic control of milling yield,dough rheology and baking quality of wheat[J].TheoreticalandAppliedGenetics,2006,112(8):1487-1495.

[18]HE X Y,ZHANG Y L,HE Z H,etal.Characterization of phytoene synthase 1 gene (Psy1) located on common wheat chromosome 7A and development of a functional marker[J].TheoreticalandAppliedGenetics,2008,116(2):213-221.

[19]PARKER G D,CHALMERS K J,RATHJEN A J,etal.Mapping loci associated with floor color in wheat (TriticumaestivumL)[J].TheoreticalandAppliedGenetics,1998,97:238-245.

[20]高鳳梅,邵立剛,王巖,等.黑龍江省春小麥主要品質(zhì)性狀和慢銹抗病基因的分子標(biāo)記檢測(cè)[J].麥類作物學(xué)報(bào),2013,33(2):243-248.

GAO F M,SHAO L G,WANG Y,etal.Molecular marker characterization of quality and disease resistant genes in Heilongjiang spring wheat varieties [J].JournalofTriticeaeCrops,2013,33(2):243-248(in Chinese with English abstract).

[21]蘆靜,胡鈺,周安定,等.新疆小麥黃色素含量基因等位變異的分子檢測(cè)及其分布[J].麥類作物學(xué)報(bào),2012,32(2):234-239.

LU J,HU Y,ZHOU A D,etal.Molecular detection and distribution of allelic variations of the gene for yellow pigment content in Xinjiang wheat cultivars[J].JournalofTriticeaeCrops,2012,32(2):234-239(in Chinese with English abstract).

[22]王憲國(guó),張鈺玉,白升升,等.寧夏小麥黃色素含量相關(guān)基因的組成與分布[J].麥類作物學(xué)報(bào),2014,34(1):8-12.

WANG X G,ZHANG Y Y,BAI SH SH,etal.Analysis on composition and distribution of genes for yellow pigment content in Ningxia wheat [J].JournalofTriticeaeCrops,2014,34(1):8-12(in Chinese with English abstract).

[23]胡鳳靈,何中虎,葛建貴,等.冬小麥品種高低分子量麥谷蛋白亞基的分子標(biāo)記檢測(cè)[J].麥類作物學(xué)報(bào),2011,31(6):1071-1076.

HU F L,HE ZH H,GE J G,etal.Characterization of high-and low-molecular weight glutenin subunits in Chinese winter wheat varieties using molecular markers[J].JournalofTriticeaeCrops,2011,31(6):1071-1076(in Chinese with English abstract).

[24]JIMENEZ M,DUBCOVSKY J.Chromosome location of genes affecting polyphenol oxidase activity in seeds of common and durum wheat[J].PlantBreeding,1999,118(5):395-398.

[25]RAMAN R,RAMAN H,JOHNSTONE K,etal.Genetic and in silico comparative mapping of the polyphenol oxidase gene in bread wheat (TriticumaestivumL.)[J].Functional&IntegrativeGenomics,2005,5(4):185-200．

[26]MA W,ZHANG W,GALE K R.Multiplex-PCR typing of high molecular weight glutenin alleles in wheat[J].Euphytica,2003,134(1):51-60.

[27]LEI Z S,GALE K R,HE Z H,etal.Y-type gene specific markers for enhanced discrimination of high-molecular weight glutenin alleles at the Glu-B1 locus in hexaploid wheat [J].JournalofCerealScience,2006,43:94-101.

[28]American Association of Cereal Chemists.Approved Methods Committee.Approved Methods of the American Association of Cereal Chemists 9th Ed.[M].St Paul Minn:American Association of Cereal Chemists,1995.

[29]胡鳳靈,何中虎,葛建貴,等.小麥品種黃色素含量和多酚氧化酶活性基因的分子標(biāo)記檢測(cè)[J].麥類作物學(xué)報(bào),2011,31(1):47-53.

HU F L,HE ZH H,GE J G,etal.Identification of genes for yellow pigment content and polyphenol oxidase activity in common wheat using molecular markers [J].JournalofTriticeaeCrops,2011,31(1):47-53(in Chinese with English abstract).

[30]HE Z H,YANG J,ZHANG Y,etal.Pan bread and dry white Chinese noodle quality in Chinese winter wheats[J].Euphytica,2004,139(3):257-267.

[31]劉麗,閻俊,張艷,等.冬播麥區(qū)Glu-1和Glu-3位點(diǎn)變異及1B/1R易位與小麥加工品質(zhì)性狀的關(guān)系[J].中國(guó)農(nóng)業(yè)科學(xué),2005,38(10):1944-1950.

LIU L,YAN J,ZHANG Y,etal.Allelic variation at the Glu-1 and Glu-3 loci and presence of 1B/1R translocation,and their effects on processin rocessing quality in cultivars and advanced lines from autumn-wown wheat regions in China [J].ScientiaAgriculturaSinica,2005,38(10):1944-1950(in Chinese with English abstract).

[32]蘆靜,何中虎,夏先春,等.新疆小麥品種高分子量谷蛋白亞基及相關(guān)品質(zhì)基因的分子標(biāo)記檢測(cè)[J].作物學(xué)報(bào),2009,35(4):647-661.

LU J,HE ZH H,XIA X CH,etal.Characterization of Xinjiang local and intoduced wheat germplasm for high molecular weight glutenin subunits and quality-related genes with molecular markers[J].ActaAgronomicaSinica,2009,35(4):647-661(in Chinese with English abstract).

[33]蘆靜,李建疆,梁曉東,等.新疆部分春小麥品種(系)相關(guān)品質(zhì)性狀基因等位變異的分子檢測(cè)[J].新疆農(nóng)業(yè)科學(xué),2010,47(12):2333-2338.

LU J,LI J J,LIANG X D,etal.Molecular detection of allelic variations of related quality traits from Xinjiang partial spring wheat cultivars [J].XinjiangAgriculturalSciences,2010,47(12):2333-2338(in Chinese with English abstract).

[34]肖永貴,劉新倫,何勝美,等.1BL·1RS易位系對(duì)小麥籽粒性狀的遺傳效應(yīng)分析[J].西北農(nóng)林科技大學(xué)學(xué)報(bào)(自然科學(xué)版),2011,39(5):140-146.

XIAO Y G,LIU X L,HE SH M,etal.Analyses of genetic effect of 1BL·1RS translocation on kernel traits in common wheat[J].JournalofNorthwestA&FUniversity(NaturalSicenceEdition),2011,39(5):140-146(in Chinese with English abstract).

Received 2015-01-05Returned2015-04-30

Foundation itemThe National Natural Science Foundation of China (No.31560390); Special Fund for Agro-scientific Research in the Public Interest(No.201503125-1);the Wheat Engineering Research Center of Gansu Province (No.144JTGA230); the National Agriculture Science Technology Achievement Transformation Fund (No.2014GB2G100140).

First authorLIU Na,female,assistant researcher.Research area:molecular and conventional breeding of wheat.E-mail:592905658@qq.com

(責(zé)任編輯：成敏Responsible editor:CHENG Min)

Molecular Marker Characterization of Main Quality Genes in 104 Gansu Wheat Varieties

LIU Na1, CAO Dong2, WANG Shihong1and YANG Wenxiong1

(1.Wheat Institute, Gansu Academy of Agricultural Sciences, Lanzhou730070, China;2.College of Agronomy, Gansu Agricultural University, Lanzhou730070, China)

AbstractSubunits of high molecular weight glutenin, yellow pigment mass fraction and 1B/1R translocation have main impacts on wheat processing quality.In this study, locus specific molecular markers for high molecular mass glutenin subunits of Dx5, Bx7, By8 and By9, PSY, 1B/1R translocation genes were used to characterize 104 wheat varieties to know the distribution of these genes in wheat varieties of Gansu province, and provide parent materials in molecular marker assisted breeding program for wheat with good quality.The results showed that, the frequency of cultivars with genes of Dx5, Bx7, By8, By9 and 1B/1R translocation were 63.46%, 76.92%, 52.88%, 84.62%,45.19% and 45.19%, respectively.The proportion of the allelic variations of Psy-A1a and Psy-A1b in these varieties were 60.58% and 21.15% respectively.Significant differences in yellow pigment mass fraction were detected between the genotypes with Psy-A1a and those with Psy-A1b (P<0.05).Generally, the frequency of single good quality gene for processing traditional food, such as steamed bread and flour, was high, the combination of good quality genes were low and the number of cultivars with high yellow pigment mass fraction was high in wheat cultivars released in Gansu province.

Key wordsGansu wheat; High molecular mass glutenin subunits; Quality genes; Molecular marker

Corresponding authorYANG Wenxiong,male,researcher.Research area:wheat breeding.E-mail:ywxm822@126.com

中圖分類號(hào)S512.1+2

文獻(xiàn)標(biāo)志碼A

文章編號(hào)1004-1389(2016)03-0353-08

通信作者：楊文雄，男，研究員，主要從事小麥育種研究。E-mail:ywxm822@126.com

基金項(xiàng)目：國(guó)家自然科學(xué)基金(31560390)；公益性行業(yè)(農(nóng)業(yè))科研專項(xiàng)(201503125-1)；甘肅省小麥工程技術(shù)研究中心專項(xiàng)(144JTGA230)；國(guó)家農(nóng)業(yè)科技成果轉(zhuǎn)化資金(2014GB2G100140)。

收稿日期：2015-01-05修回日期：2015-04-30

網(wǎng)絡(luò)出版日期：2016-03-06

網(wǎng)絡(luò)出版地址：http://www.cnki.net/kcms/detail/61.1220.S.20160306.1610.012.html

第一作者：柳娜，女，助理研究員，從事小麥分子和常規(guī)育種研究。E-mail:592905658@qq.com