陕西小麦籽粒硬度及其基因型分析

籽粒硬度与小麦市场分级定价、磨粉品质和食品加工品质密切相关。为给小麦品种选育和品种推广提供参考依据,用单籽粒谷物硬度测试仪测定了169份陕西小麦品种（系）的籽粒硬度,并利用分子标记检测和基因序列分析确定了硬质麦的基因组成。硬度测定结果表明,陕西参试小麦品种（系）存在硬质麦、混合麦和软质麦3种类型,分别为121、11和37份,依次占71.6%、6.5%和21.9%。陕西不同地区3种籽粒硬度类型所占比例明显不同。基因型分析结果表明,陕西硬质麦存在4种基因型,即PinaD1b、PinbD1b、PinbD1d和PinbD1p,分别有14、97、2和8份材料,占硬质麦比例依次为11.6%、80.2%、1.6%和6.6%。总体而言,陕西小麦以硬质麦为主,硬质麦主要由PinbD1b基因型组成。     

长江中下游麦区小麦品种籽粒硬度及puroindoline基因等位变异的分子检测

为了明确长江中下游麦区小麦籽粒硬度及puroindoline基因型的分布,以该麦区105份小麦育成品种为材料,利用单籽粒硬度仪(SKCS)测定其籽粒硬度,利用分子标记检测和基因序列分析鉴定puroindoline基因的等位变异。结果表明,在长江中下游麦区历年育成的小麦品种中软质麦比例较高,占52.4%,硬质麦和混合麦分别占38.1%和9.5%;硬质麦和混合麦中存在Pinb-D1b、Pina-D1b和Pinb-D1p三种变异类型,突变频率分别为29.5%、10.5%和3.8%。     

CIMMYT普通冬小麦品种的籽粒硬度及Puroindoline基因等位变异

为给中国小麦种质资源引进、利用和品质改良提供信息,以国际玉米小麦改良中心（CIMMYT）土耳其育种站提供的192份普通冬小麦新品系为材料,采用单籽粒谷物特性测试仪、特异引物PCR扩增和改进的SDS-PAGE凝胶电泳方法对其SKCS硬度及Puroindoline基因型进行了鉴定和分析。结果表明,CIMMYT普通冬小麦以硬质类型为主,但SKCS硬度值普遍偏低,平均值仅为60．7。所调查的192份材料中,硬质麦119份,占62．0％;软质麦49份,占25．5％;混合麦24份,占12．5％。硬质小麦共有4种基因型,分别为PinA蛋白缺失类型（Pina-D1b／P1nb-D1a）90份、Pina-D1a／Pinb—D1b类型27份Pina—D1a／Pinb—D1d类型2份和Pina-D1b/Pinb-D1d类型1份,以PinA蛋白缺失类型为主,占总数的75．6％。Pina—D1b／Pinb-D1d为Pina和Pinb基因的双突变类型。CIMMYT普通冬小麦籽粒硬度及其Puroindoline基因变异类型和分布的信息能够为中国冬小麦品质改良提供理论依据。     

45份青海春小麦品种矮秆基因检测及效应分析

为明确矮秆基因Rht-B1和Rht-D1在青海春麦区小麦品种中的分布，以青海春麦区历年种植的45份春小麦品种（25份育成品种，14份引进品种，6份地方品种）为材料，利用KASP标记技术检测其两个矮秆基因的分布，并分析这两个矮秆基因对株高、穗长、小穗数和穗粒数的影响。结果表明，Rht-B1位点上存在Rht-B1a和Rht-B1b两种等位变异，分布频率分别为91.11%和8.89%；Rht-D1位点上存在Rht-D1a和Rht-D1b两种等位变异，分布频率分别为86.67%和13.33%。25份育成品种中，Rht-B1b和Rht-D1b的分布频率分别为16.00%和12.00%，未发现Rht-B1a和Rht-D1a；引进品种中仅含有Rht-D1b，分布频率为21.43%；地方品种中未发现Rht-B1b和Rht-D1b。Rht-B1b和Rht-D1b均具有一定的降秆效应，Rht-D1b的降秆效应大于Rht-B1b。携带Rht-B1a/Rht-D1b品种的株高显著低于携带Rht-B1a/Rht-D1a和Rht-B1b/Rht-D1a的品种，该组合类型不仅可使株高降低，也可增加小穗数和穗粒数；未发现携带Rht-B1b/Rht-D1b组合类型的品种。     

新疆小麦品种Wx基因组成及其淀粉糊化特性研究

为了给新疆小麦淀粉品质改良和优质面条小麦新品种选育提供理论依据,利用WxA1、WxB1和WxD1位点的3个STS标记（MAG264、BDFLBRD和MAG269）对250份新疆小麦品种（包括157份冬小麦品种和93份春小麦品种）Wx基因的组成进行了鉴定,同时测试了185份冬、春小麦品种的淀粉糊化特性。结果表明,新疆小麦品种（系）Wx基因组成比较单一,以野生型（WxA1a/WxB1a/WxD1a）为主,占参试品种总数的84.0%,除了在WxB1位点发现亚基缺失的品种（缺失比例为13.6%）外,没有发现其他亚基缺失类型。WxB1蛋白亚基缺失比例在冬、春小麦间差异较大,春小麦品种WxB1亚基缺失比例（21.5%）要高于冬小麦品种（8.9%）;农家品种、引进品种和育成品种中WxB1亚基缺失比例也各不相同,依次为0%、5.6%和22.6%。分析表明,新疆小麦品种淀粉糊化特性变异范围相当广泛,冬、春小麦品种在峰值黏度、稀澥值和反弹值等主要淀粉指标上均达到极显著差异,春小麦品种的淀粉特性优于冬小麦品种,并且WxB1亚基缺失体淀粉糊化特性的平均表现总体优于非缺失体。本研究所用3个分子标记对小麦基因组DNA扩增的稳定性、重复性较好,检测结果准确、可靠,可以用于小麦品种Wx基因组成的鉴定和糯质专用小麦的分子标记辅助选育。     

新疆小麦品种籽粒硬度及puroindoline基因等位变异的分子检测

为了解新疆小麦品种籽粒硬度概况和puroindoline基因等位变异类型及其分布,以121份新疆冬、春小麦品种(包括51份农家品种和70份育成品种)为材料,采用单粒谷物特性测试仪(SKCS)和分子标记技术,对其SKCS硬度及puroindoline基因型进行了测试和鉴定。结果表明,新疆小麦硬度变化范围较大,以硬质麦为主,占61.2%。冬性农家品种籽粒硬度高于春性农家品种,冬、春小麦育成品种籽粒硬度基本相同(60.6和60.8)。新疆小麦puroindoline基因的类型丰富,共检测到野生型(Pinb-D1 a)、Pina-D1 b、Pinb-D1 b、Pinb-D1 p、Pinb-D1 ab和Pinb-D1 ac6种类型。硬质麦以Pina-D1 b、Pinb-D1 p、Pinb-D1 b3种突变类型为主,其频率分别为33.8%、31.1%和28.4%。冬性农家品种含有3种类型,以Pina-D1 a/Pinb-D1 b类型居多,春性农家品种含有4种类型,以野生型(Pina-D1 a/Pinb-D1 a)为主,冬、春麦育成品种含有4种常见类型,其中冬麦以Pinb-D1 b类型为主,其频率为39.5%,春麦以Pina-D1 b(PINA缺失)类型为主,其频率为59.4%。另外,冬性农家品种中有4个品种属于Pina-D1 a/Pinb-D1 ab类型,春性农家品种中有1个品种属于Pina-D1 a/Pinb-D1 ac类型,均属硬质麦的稀有突变类型。不同puroindoline基因型的籽粒硬度大小也存在差异,其中Pinb-D1 ab突变型的硬度值最高,Pinb-D1 a最低,并且Pina-D1 b、Pinb-D1 b和Pinb-D1 p3种硬质类型的籽粒硬度没有显著性差异。     

Pins基因等位变异对磨粉品质和新疆拉面加工品质的影响

为了探讨新疆冬小麦品种Pins基因等位变异对小麦磨粉品质和新疆拉面加工品质的影响,对109份新疆冬小麦品种的籽粒硬度及其Pins基因等位变异、磨粉品质和新疆拉面加工品质进行测定,初步分析了新疆冬小麦品种资源籽粒硬度Pins基因的分布规律以及不同 Pins基因等位变异对籽粒硬度、磨粉品质和新疆拉面加工品质的影响。结果表明,新疆冬小麦品种属硬质麦类型,Pins基因型以 Pina-D1a、 Pinb-D1b和 Pina-D1a/ Pinb-D1b为主, Pins突变类型及Pins突变基因型组合类型小麦的籽粒硬度均显著高于野生型, Pinb-D1a基因型小麦的籽粒硬度最低,L^*值和a^*值最高,b^*值最低; Pinb-D1ab基因型小麦的吸水率最高。不同Pins基因型组合中,野生型小麦的籽粒硬度、b^*值和吸水率最低; Pina-D1a/ Pinb-D1aa的出粉率最高, Pina-D1a/ Pinb-D1ab的灰分含量最低,吸水率最高。Pins基因及其基因型组合对新疆拉面加工品质无直接影响,主要通过对灰分、面粉色泽和吸水率等磨粉品质的作用对新疆拉面产生间接影响。优质新疆拉面品种中,Pinb基因突变对新疆拉面加工品质的影响大于Pina基因突变,育种中应优先选择Pinb 基因突变型材料,其中 Pina-D1a/ Pinb-D1b可以作为重点选择的基因型组合。     

CIMMYT普通冬小麦品种的籽粒硬度及Puroindoline基因等位变异

为给中国小麦种质资源引进、利用和品质改良提供信息,以国际玉米小麦改良中心(CIMMYT)土耳其育种站提供的192份普通冬小麦新品系为材料,采用单籽粒谷物特性测试仪、特异引物PCR扩增和改进的SDS-PAGE凝胶电泳方法对其SKCS硬度及Puroindoline基因型进行了鉴定和分析.结果表明,CIMMYT普通冬小麦以硬质类型为主,但SKCS硬度值普遍偏低,平均值仅为60.7.所调查的192份材料中,硬质麦119份,占62.0%;软质麦49份,占25.5%;混合麦24份,占12.5%.硬质小麦共有4种基因型,分别为PinA蛋白缺失类型(Pina-D1b/Pinb-D1a)90份、Pina-D1a/Pinb-D1b类型27份、Pina-D1a/Pinb-D1d类型2份和Pina D1b/Pinb-D1d类型1份,以PinA蛋白缺失类型为主,占总数的75.6%.Pina-D1b/Pinb-D1d为Pina和Pinb基因的双突变类型.CIMMYT普通冬小麦籽粒硬度及其Puroindoline基因变异类型和分布的信息能够为中国冬小麦品质改良提供理论依据.     

黄淮麦区小麦品种和CIMMYT材料的矮秆基因型及其对株高和胚芽鞘的影响

为促进矮秆基因在小麦品种遗传改良中的应用,以131份黄淮麦区小麦品种和31份CIMMYT（国际玉米小麦改良中心）小麦材料为研究对象,用分子标记检测 Rht-B1a、 Rht-B1b、 Rht-D1a、 Rht-D1b和 Rht8等小麦矮秆基因,并结合赤霉素处理,分析了供试材料所含矮秆基因类型及其对株高和胚芽鞘长度的影响。结果表明,供试材料中,63份含有矮秆基因 Rht-B1b,66份含有矮秆基因 Rht-D1b,105份含有矮秆基因 Rht8,分别占供试材料的38.88%、40.74%和64.81%。黄淮麦区小麦品种大多数含有矮秆基因 Rht-D1b,CIMMYT小麦中大多数含有矮秆基因 Rht-B1b。赤霉素处理结果显示,供试小麦材料中,有52份对赤霉素敏感,占被检测材料的32.09%。该批材料中,矮秆基因 Rht-D1b和 Rht8的降秆效应最大, Rht-B1b和 Rht-D1b的降秆效应分别是14.69%和17.74%,各基因组合的降秆效应表现为 Rht-D1b+ Rht8＞ Rht-D1b＞ Rht-B1b+ Rht8＞ Rht-B1b＞ Rht8。同时含有矮秆基因 Rht-B1b和 Rht-D1b的材料缩短胚芽鞘长度的效应最强,缩短效应为26.20%,缩短胚芽鞘长度的效应表现为 Rht-B1b+ Rht-D1b＞ Rht-B1b+ Rht-D1b + Rht8＞ Rht-D1b＞ Rht-D1b+ Rht8＞ Rht-B1b＞ Rht-B1b+ Rht8＞ Rht8。基于以上结果,矮秆基因 Rht8能够在降低小麦材料株高的同时不影响胚芽鞘长度,因此应重视其在矮化育种中的应用。     

不同高分子量麦谷蛋白亚基小麦品种（系）的粒重增长特性分析

为给小麦高产优质育种提供参考,以黄淮麦区35个主栽小麦品种（系）为材料,利用SDSPAGE电泳技术对其进行了高分子量麦谷蛋白亚基鉴定,并对各参试品种的灌浆特性进行了分析。结果表明,35个黄淮麦区小麦品种（系）中 GluA1、GluB1和 GluD1位点分别只有2、5和4种亚基类型,分别以Null、7+9和2+12亚基为主,所占比例分别为71.4%、45.7%和62.7%。依据 GluB1和 GluD1位点的亚基特点,将参试品种（系）分为三类。对这三类品种（系）的灌浆期粒重增长特性分析发现,第I类（亚基组合：7+8/2+12）品种（系）灌浆速率较快,最终粒重较高;第II类（亚基组合：7+9/2+12）品种灌浆持续时间较长,但灌浆速率不及第I类,最终粒重较高;第III类（亚基组合：7/5+10或7+8/5+10）品种（系）灌浆持续时间较短,且灌浆速率较慢,最终粒重不高。三类品种（系）相比,粒重增长速率在灌浆前期基本相同,但35 d以后,第III类品种（系）粒重增长速率明显减慢,这可能是导致其最终粒重显著低于前两类品种（系）的主要原因。     

Mapping kernel texture in a soft durum (Triticum turgidum subsp. durum) wheat population

Kernel texture (”hardness”) is one of the major determinants of wheat quality and is primarily controlled by the Puroindoline (Pin) genes, located at the Hardness (Ha) locus. The absence of the Ha locus is responsible for the extremely hard kernels of durum wheat (T. turgidum subsp. durum). Recently, the Pin genes from a soft common wheat variety were introgressed into durum wheat through homoeologous recombination. The objective of this study was to map kernel hardness in a soft durum wheat population derived from the cross between the varieties “Creso” and “Langdon 1–678“. In all, 428 F₆ lines were evaluated for kernel hardness through the Single Kernel Characterization System; Hardness Index (HI) values ranged from −2 to 44. The same lines were genotyped using genotyping-by-sequencing, targeted amplicon sequencing, and sequence-tagged-site markers. A total of 8537 markers were used to conduct single marker-trait association analysis and two major significant regions were identified on chromosomes 3AL and 6AS each responsible for an additive effect of ∼6 HI units. Kompetitive allele specific markers targeting these regions were selected and tested in the whole population. To date, this is the first study to investigate the genetic factors behind hardness variation in durum wheat.     

The influence of temperature extremes on some quality and starch characteristics in bread,biscuit and durum wheat

Environmental conditions during grain-fill can affect the duration of protein accumulation and starch deposition, and thus play an important role in grain yield and flour quality of wheat. Two bread-, one durum- and one biscuit wheat were exposed to extreme low (−5.5 °C for 3 h) and high (32 °C/15 °C day/night for three days) temperatures during grain filling under controlled conditions for two consecutive seasons. Flour protein content was increased significantly in one bread wheat, Kariega, under heat stress. Cold stress significantly reduced SDS sedimentation in both bread wheats. Kernel weight and diameter were significantly decreased at both stress treatments for the two bread wheats. Kernel characteristics of the biscuit wheat were thermo stable. Kernel hardness was reduced in the durum wheat for the heat treatment. Durum wheat had consistently low SDS sedimentation values and the bread wheat high values. Across the two seasons, the starch content in one bread wheat was significantly reduced by both high and low temperatures, as is reflected in the reduction of weight and diameter of these kernels. In the durum wheat, only heat caused a significant reduction in starch content, which is again reflected in the reduction of kernel weight and diameter.     

小麦籽粒发育时期Puroindolines蛋白与硬度的关系

为探讨Puroindolines蛋白的表达特点与籽粒硬度的关系,采用改进的SDS-PAGE凝胶分析了不同硬度小麦品种的籽粒在各个发育时期Puroindolines蛋白的表达.结果表明,不同硬度的小麦籽粒中总Puroindolines(PinA和PinB)蛋白的表达量差异不大,但与胚乳淀粉颗粒结合的Puroindolines蛋白量差异非常明显:在籽粒发育的不同时期,软质小麦籽粒淀粉粒表面结合的Puroindolines蛋白量显著高于硬质小麦;基因型同为野生型但硬度有差异的品种,籽粒较软的材料其淀粉粒表面结合的Puroindolines蛋白量也明显高于较硬的材料,说明该蛋白的结合特性是决定籽粒硬度的直接原因.结果还表明,胚乳中水溶性戊聚糖与籽粒硬度关系密切.     

Transformation of Pinb-D1x to soft wheat produces hard wheat kernel texture

Kernel hardness - a key quality trait of common wheat (Triticum aestivum L.) - is mainly conditioned by the Pina and Pinb genes. Mutation or deletion of Pina or Pinb increases kernel hardness, resulting in a hard wheat kernel texture. Here, Pinb-D1x gene was cloned from a hard wheat landrace Kashibaipi and transformed into a soft wheat cultivar Yangmai19 to assess its effect on kernel hardness and flour properties. PCR, RT-PCR and Western blot data confirmed the successful transformation and overexpression of Pinb-D1x gene in transgenic offsprings. The data of single kernel characterization system and scanning electron microscopy revealed that the introduction of Pinb-D1x in soft wheat increased the kernel hardness significantly and changed the internal structure of the kernel. Similarly, transgenic lines exhibited hard wheat like flour properties; flour whiteness and pasting temperature were significantly reduced in the transgenic lines, while the total protein content, damaged starch content, and compound parameter in the Mixograph tests (PT × TW value) showed a significant increase over the wildtype. The results showed that the transformation of the Pinb variants is a powerful strategy to alter the kernel hardness and flour properties in wheat breeding.     

不同硬度小麦品质差异的分析

为了解不同硬度级别小麦的品质差异,并为新国标<小麦硬度测定硬度指数法>在中国小麦硬度测定中的推广和应用提供参考依据.选取硬质小麦、中硬小麦和软质小麦各6个品种,分别对其品质性状进行测试.结果表明,小麦耔粒硬度与沉淀值、湿面筋含量、破损淀粉含量、面包评分等小麦品质特性和加工特性达极显著相关关系.容重、出粉率和破损淀粉含量随着硬度指数值的升高逐渐增加,但其差异仅在软质小麦与硬质小麦之间达显著水平.硬质小麦与中硬小麦之间未达显著水平.随着硬度指数的升高,面粉蛋白质含量逐渐升高,粗淀粉含量逐渐降低,且在不同硬度小麦级别之间都迭显著水平;直链淀粉占样品的比重逐渐降低,支链淀粉含量升高;面包、面条的加工品质逐步得到了改善,其中面包品质的改善较面条品质更为显著.     

小麦籽粒硬度研究进展

小麦籽粒硬度是当前小麦育种的主要目标之一,对小麦的制粉品质有重要影响,是小麦品质评价的重要指标之一。本文从小麦籽粒硬度测定方法、籽粒硬度与胚乳结构的关系、籽粒硬度对面粉品质和面制品品质的影响、籽粒硬度的遗传规律、品种与环境对籽粒硬度的影响、籽粒硬度育种改良等方面介绍了国内外相关领域的研究现状,并对未来发展提出展望,为小麦育种中籽粒硬度的改良提供参考。     

Analysis of Pina and Pinb alleles in the micro-core collections of Chinese wheat germplasm by Ecotilling and identification of a novel Pinb allele

Kernel hardness is mainly conditioned by allelic variations of Pina-D1 and Pinb-D1 genes located on the short arm of chromosome 5D. In this work, the Ecotilling approach was optimized to investigate Pina and Pinb alleles in the micro-core collections of Chinese wheat germplasm, and three Pina and eight Pinb alleles were found. Generally, more Pinb alleles were detected in the accessions coming from the regions that grow winter or a mixture of spring and winter wheats. This was particularly evident for the Southwestern winter wheat, Xinjiang winter–spring wheat and Yellow and Huai River Valley winter wheat regions. A novel variant (designated as Pinb-D1x) was discovered in one of the accessions from the Xinjiang winter–spring wheat region. Compared to wild type (WT) allele Pinb-D1a, two nucleotide substitutions occurred in the coding region of Pinb-D1x, one (at nucleotide position 257) resulting in the replacement of a WT cysteine residue by tyrosine and the other (at nucleotide position 382) creating a premature stop codon. The implications of our data to understanding the diversity of Pina and Pinb alleles in wheat and to future molecular breeding of wheat kernel hardness are discussed.     

A new puroindoline b mutation present in Chinese winter wheat cultivar Jingdong 11

Kernel hardness is one of the most important characteristics in determining utilization and marketing of bread wheat. Genes coding for puroindoline a and b (PINA and PINB) were located at the Ha locus and designated as Pina-D1 and Pinb-D1, respectively. The coding sequence of the Pinb gene in a Chinese winter wheat cultivar Jingdong 11 (Triticum aestivum L.) was amplified with polymerase chain reaction (PCR), and the obtained 447-bp fragment sequenced from two strands, and compared with the eight known Pinb alleles. The results showed that Jingdong 11 possessed a new Pinb allele not reported previously, and was designated as Pinb-D1q. It is characterized by a single base T to G substitution, which results in a tryptophan to leucine substitution (TGG to TTG) at position 44 and is most likely the cause of hard grain texture in Jingdong 11. The characterization of Pinb-D1 alleles would be helpful in manipulating grain hardness of bread wheat in breeding programs.