水稻粒长基因GL3的遗传分析和分子标记定位

为了解析水稻粒长的遗传机制,以大粒水稻品种‘80018-TR161-2-1’和小粒水稻品种‘日本小黑稻’及其F2代200个株系和F2：3家系为材料,分析水稻粒长的遗传学性状。结果表明,谷粒长度的分离比在F2及F2：3家系中都表现为3：1,长粒性状受1对隐性核基因控制,命名为GL3。用简单重复序列（simple sequence repeat,ssR）分子标记结合群体分组混合分析的方法,将此种基因定位在水稻第3号染色体上SSR标记PSM379和RM16之间,它们的遗传距离分别为4．0cM和11．2cM。     

一份新型水稻极度分蘖突变体的遗传分析及分子标记定位

在三系杂交水稻保持系绵香1B（M1B）和一个雄性不育材料GMS-1的杂交后代中发现一株极度分蘖突变体（命名为ext．M1B）,其分蘖数为121。对ext-M1B与5个正常分蘖水稻品种杂交F1和F2代的遗传分析表明,ext-M1B的极度分蘖特性受一对隐性核基因控制。以2480B／ext-M1B的F2代作定位群体,用分子标记将ext-M1B的突变基因定位于水稻第6染色体短臂,该基因与微卫星标记RM197、RM584和RM225的遗传距离分别为3．8cM、5．1cM和5．2cM,认为ext-M1B突变基因是一个新的水稻极度分蘖基因,暂命名为ext-M1B（t）。     

一个水稻显性高秆突变体的遗传分析和基因定位

从水稻(Oryza sativa L.)的两个半矮秆籼稻品种6442S-7和蜀恢881杂交F2代群体中发现一个高秆突变体D111,其株高和秆长分别比亲本蜀恢881增加63.0%和87.0%.用205个微卫星标记分析D¨1及其原始亲本6442S-7和蜀恢881之间的基因组DNA多态性,结果未发现D111具有2个原始亲本都没有的新带型,证明D1¨的确是6442S-7和蜀恢881的杂交后代发生基因突变产生的.将D111分别与蜀恢881、蜀恢527、明恢63、9311、IR68、G46B等6个半矮秆品种和高秆对照品种南京6号杂交,分析F1和F2代株高的遗传行为,结果表明D1¨的高秆性状由一对显性基因控制,且该基因与南京6号的高秆基因紧密连锁或等位.以蜀恢527/D111 F2群体为定位群体,运用微卫星标记将D111显性高秆突变基因定位于水稻第一染色体长臂,与RM212、RM302和RM472的遗传距离分别是27.7 cM、25.5 cM和6.0 cM,该基因暂命名为LC(t).认为D111是首例从半矮秆品种自然突变产生的水稻显性高秆突变体,LC(t)为首次定位的水稻显性高秆突变基因.此外,将上述基因定位结果与Causse等(1994)和Temnykh等(2000,2001)发表的水稻分子连锁图谱进行比较,发现LC(t)基因恰巧位于与水稻"绿色革命基因"sd1相同或十分相近的染色体区域,因此,还就LC(t)基因与sd1基因之间的可能关系进行了讨论.     

一个水稻显性高秆突变体的遗传分析和基因定位

从水稻(Oryza sativa L.)的两个半矮秆籼稻品种6442S-7和蜀恢881杂交F2代群体中发现一个高秆突变体D111,其株高和秆长分别比亲本蜀恢881增加63.0%和87.0%。用205个微卫星标记分析D111及其原始亲本6442S-7和蜀恢881之间的基因组DNA多态性,结果未发现D111具有2个原始亲本都没有的新带型,证明D111的确是6442S-7和蜀恢881的杂交后代发生基因突变产生的。将D111分别与蜀恢881、蜀恢527、明恢63、9311、IR68、G46B等6个半矮秆品种和高秆对照品种南京6号杂交,分析F1和F2代株高的遗传行为,结果表明D111的高秆性状由一对显性基因控制,且该基因与南京6号的高秆基因紧密连锁或等位。以蜀恢527/D111 F2群体为定位群体,运用微卫星标记将D111显性高秆突变基因定位于水稻第一染色体长臂,与RM212、RM302和RM472的遗传距离分别是27.7 cM、25.5 cM和6.0 cM,该基因暂命名为LC(t)。认为D111是首例从半矮秆品种自然突变产生的水稻显性高秆突变体,LC(t)为首次定位的水稻显性高秆突变基因。此外,将上述基因定位结果与Causse等(1994)和Temnykh等(2000; 2001)发表的水稻分子连锁图谱进行比较,发现LC(t)基因恰巧位于与水稻“绿色革命基因”sd1相同或十分相近的染色体区域,因此,还就LC(t)基因与sd1基因之间的可能关系进行了讨论。     

矮泰引-3中半矮秆基因的分子定位

矮泰引-3的矮生性状受两对独立遗传的半矮秆基因控制,利用SSR标记将这两个矮秆基因分别定位到第1和第4染色体上。等位性测交的结果表明,位于第1染色体上的矮秆基因与sd1是等位的,所以仍然称其为sd1;而位于第4染色体上的矮秆基因是一个新基因,暂命名为sdt2。利用SSR标记将sd1定位于RM297、RM302和RM212的同一侧,而与OSR3共分离,它们之间的位置关系可能是RM297-RM302-RM212-OSR3-sd1,遗传距离分别为4．7cM、0cM、0．8cM和0cM,这与sd1在第1染色体长臂上的确切位置是基本一致的。利用已有的SSR标记和拓展的SSR标记将sdt2定位于SSR332、RM1305和RM5633、RM307、RM401之间,它们的排列位置可能是SSR332-RM1305-sdt2-RM5633-RM307-RM401,它们之间的遗传距离分别为11．6cM、3．8cM、0．4cM、0cM和0．4cM。     

水稻顶节间长度控制基因(EUI)的精细定位

通过对一水稻顶节间特异伸长突变体Mh-1进行经典遗传学和基因定位分析,认为该表型是一个核基因隐性突变所致。利用Mh-1与正常的T65-sd1杂交的F2群体对该位点定位研究分析,发现其与水稻第5条染色体长臂STS标记E30531和CAPS标记C903连锁,遗传距离分别为6．7cM和2．8cM。经进一步发展新的分子标记,将该基因精确定位在0．3cM的区域,为进一步克隆和研究该基因的分子机制奠定基础。     

一个新的水稻小粒矮秆基因的分子标记定位及效应分析

一个新的水稻小粒矮秆基因的分子标记定位及效应分析 李秀兰 吴 成 邓晓建 王平荣 李仁端 杨志荣     

水稻叶状颖壳突变体Oslh的遗传分析和OsLH基因的定位

通过γ射线诱变,从粳稻品种9522的M2代中筛选出一株具有叶状颖壳的突变体,定名Oslh(1h=leafy hull).Oslh突变体的开花时间要比野生型晚15 d左右,内外稃和浆片发育成了叶片状器官.Oslh突变体与粳稻品种9522回交结果表明Oslh突变性状可能由单核基因隐性突变造成.以Oslh突变体与籼稻品种广陆矮4号杂交的F2代群体为基因定位群体,利用SSR和InDel分子标记将Oslh突变位点定位在3号染色体上的SSR标记RM5475和InDel标记GY305之间,遗传距离分别为2.5 cM和1.9 cM.这些结果为克隆OsLH基因和研究花器官发育的调控机理奠定了基础.     

一个控制水稻籽粒长度主效基因--Lk-4(t)的BC2F2群体定位及其遗传效应分析

水稻籽粒大小和形状是影响稻米外观品质和产量的重要影响因素,对控制这些性状基因的定位和克隆有助于弄清籽粒大小基因的表达模式和相应的代谢系统,最终实现该性状的自由调控。运用SSR和CAPs标记对来源于蜀恢527//蜀恢527／小粒回交组合BC2F2群体800隐性长粒单株进行分析,定位了一个控制水稻籽粒长短的基因,Lk-4（t）。对F2和BC2F2群体籽粒大小形状和千粒重的遗传分析表明,回交能将大部分对目的基因效应具有干扰修饰作用的微效基因多态性除去,从而有利于对目的基因型的准确鉴定;在F2和BC2F2群体中只发现两类籽粒长短表现型,即短粒和长粒,并且二者分离比例符合3：1的典型一对等位基因分离比例。这说明群体中籽粒长短变异是受一对基因控制。通过对BC2F2群体中隐性（长粒）单株进行分子标记分析,将这个控制籽粒长短的主效基因定位在3个CAPs标记,P1-EcoRV,P2-SacⅠ和P3-MboⅠ附近。连锁分析表明,Lk-4（t）位于水稻第3染色体着丝粒附近,离标记P1-EcoRⅤ和P2-SacⅠ分别有0．90cM和0．50cM的距离。     

水稻长穗颈基因eui紧密连锁SSR标记获得

02428h是从半矮秆材料02428体细胞培养后代中发现的隐性高秆突变体, 其株高性状由1对长穗颈基因eui和1对半矮秆基因sd-1共同控制。以02428h与半矮秆材料南京11杂交的F₂为作图群体, 利用Gramene公布的SSR标记和根据NCBI中的BAC序列自己新开发的SSR标记, 将eui基因定位在第5染色体上的RM3673和RM0012之间, 两侧遗传距离分别为0.3 cM和1.0 cM, 为该基因的分子标记辅助选择奠定了基础。     

甘蓝型油菜矮秆突变材料99CDAM的发现及遗传分析

随着杂种优势的利用,油菜株高增加了20cm以上,导致油菜生长后期倒伏的风险加大。通过利用特殊矮秆基因来控制株高将是解决倒伏问题的有效方法。在甘蓝型油菜自交多代品系中发现了一个株高85cm左右的矮秆突变株99CDAM,该突变株具有开花早、分枝多等优良特性,产量性状和品质性状较好,各性状都能稳定遗传,具有重要的育种价值。对99CDAM和高秆品系2091、7045和7350的正反交F1以及99CDAM和2091的F2、BC1及F2:3代的遗传分析结果表明： 99CDAM的矮秆性状受3对隐性基因控制,存在母体细胞质效应,与以往的甘蓝型矮秆油菜有明显的区别。     

青海育成小麦5个主效矮秆基因的分子检测

为系统了解青海小麦矮秆基因的分布特点,并进一步为青海高原小麦的株高育种提供优异种质资源。本研究利用5个矮秆基因的特异性分子标记对82份青海小麦品种资源中的矮秆基因进行了检测,并对不同矮秆基因的降秆效应进行了分析。结果表明:82份青海育成小麦品种中有49份材料至少含有一个矮秆基因,其中Rht-B1b的分布频率最高,约占参试材料的28.0%,其次是分布频率为23.2%的Rht8基因,而矮秆基因Rht-D1b、Rht5以及Rht12的分布频率分别为9.8%、13.4%、9.8%。在49份含有不同种类矮秆基因的材料中,其中16份材料同时含有2种及以上的矮秆基因,即RhtB1b和Rht8、Rht-D1b和Rht8、Rht-B1b和Rht5、Rht-D1b和Rht5、Rht8和Rht5、Rht-B1b和Rht12、Rht5和Rht12,并未发现同时含有矮秆基因Rht-B1b和Rht-D1b的品种;2份材料分别含有3种矮秆基因,即Rht-B1b、Rht8、Rht12和Rht-B1b、Rht5、Rht8;其余31份材料仅含有1种矮秆基因。82份青海育成小麦材料中仅含有Rht-B1b的材料11份,平均株高为86.2 cm,其降秆效应为5.7%;只含有Rht-D1b的材料有5份,平均株高为84.9 cm,其降秆效应为7.1%;仅含有Rht8的材料有9份,平均株高为88.6 cm,其降秆效应为3.1%。因此,在青海育成小麦品种中,矮秆基因的降秆效应为Rht-D1bRht-B1bRht8。     

Contrasting Characteristics of Lodging Resistance in Two Super-Rice Hybrids Differing in Harvest Index

The “super rice” breeding program in China has been successful in developing high-yielding hybrids, including few with high harvest index values. However, there is limited information on the relationship between lodging resistance and harvest index, and the mechanisms underlying the relationship in super-rice hybrids. In this study, a two-year field experiment was conducted to compare lodging resistance and its related traits between two super-rice hybrids differing in harvest index, i.e., Guiliangyou 2 (G2) with a high harvest index and Y-liangyou 1 (Y1) with a typical harvest index of modern high-yielding rice varieties. Results showed that compared to Y1, G2 was lower in plant height due to its lower aboveground N uptake, and its higher stem breaking resistance (i.e., lower stem breaking index) resulted from a lower stem height at its center of gravity. Consequently, G2 had a higher lodging resistance (i.e., lower plant lodging index) than Y1. This study suggests that developing super-rice hybrids with high harvest index values is a possible way to achieve both high grain yield and strong lodging resistance in rice.     

粳稻矮秆品种苗期和成株期对赤霉素反应的研究

本试验对28个粳稻矮秆品种对GA3的反应进行了研究,结果表明粳稻矮秆品种对GA3反应较为复杂,苗期与成株期反应,大多数品种表现为趋势基本一致,但也存在着其它种类型表现;在粳稻中,对GA3反应敏感的矮秆品种并不一定携带含有sd-1等位基因,但含有sd-1基因的品种表现为敏感,不含有sd-1基因的表现为低敏感或不敏感,据此可以初步确定矮4和紫叶矮含有与sd-1非等位的矮秆基因,是一种新的矮源;GA3对株高构成因素的影响表现因品种不同而异,但以基部一、二节间为主.     

应用SSR分子标记比较佳禾早占水稻组培苗与种植苗的性状差异

株高是水稻重要的农艺形状之一,植株过高将导致倒伏和减产,目前,很多新的技术究被用来鉴定,图位克隆与水稻株高相关的基因及机理的研究,本实验选择优质早籼稻佳禾早占种植苗和经过组培获得的矮化突变水稻为材料,为研究比较它们间的遗传物质上差异,根据康耐尔大学的资料设计了311对SSR引物对佳禾早占种植材料和组培材料进行分析,对两种材料进行PCR多态性扩增,结果发现两者间存在多态性的引物有88对,多态性比例达到30.3%。在矮杆材料中不但验证了已报道的11个与调控株高性状基因相连锁的标记连锁群,同时在第3号染色体和第9号染色体上还获得了两个以前基本未有报道的标记集中分布区域。结果说明,该培养基培育出的佳禾早占水稻后代所表现出的矮杆性状与亲本在遗传物质上确有明显差别。该结果有助于挖掘和定位新的矮杆基因,并有利于今后在水稻育种中进行水稻株高性状的控制,同时也为开展矮化性状机理的研究提供有利的实验材料。     

Candidacy of a chitin-inducible gibberellin-responsive gene for a major locus affecting plant height in rice that is closely linked to Green Revolution gene <Emphasis Type="Italic">sd1</Emphasis>

Appropriate plant height is crucial for lodging resistance to improve the rice crop yield. The application of semi-dwarf 1 led to the green revolution in the 1960s, by predominantly increasing the rice yield. However, the frequent use of single sd1 gene sources may cause genetic vulnerability to pests and diseases. Identifying useful novel semi-dwarf genes is important for the genetic manipulation of plant architecture in practical rice breeding. In this study, introgression lines derived from two parents contrasting in plant height, Zhenshan 97 and Pokkali were employed to locate a gene with a large effect on plant height by the bulk segregant analysis method. A major gene, ph1, was mapped to a region closely linked to sd1 on chromosome 1; the additive effects of ph1 were more than 50 cm on the plant height and 2 days on the heading date in a BC₄F₂ population and its progeny. ph1 was then fine mapped to BAC AP003227. Gene annotation indicated that LOC_OS01g65990 encoding a chitin-inducible gibberellin-responsive protein (CIGR), which belongs to the GRAS family, might be the right candidate gene of ph1. Co-segregation analysis of the candidate gene-derived marker finally confirmed its identity as the candidate gene. A higher expression level of the CIGR was detected in all the tested tissues in tall plants compared to those of short plants, especially in the young leaf sheath containing elongating tissues, which indicated its importance role in regulating plant height. ph1 showed a tremendous genetic effect on plant height, which is distinct from sd1 and could be a new resource for breeding semi-dwarf varieties.     

Improvement of lodging resistance with QTLs for stem diameter in rice (Oryza sativa L.)

Varietal differences among ten rice cultivars showed that stem diameter is a key factor in lodging resistance (measured in terms of pushing resistance). Two near-isogenic lines (NILs) were selected from a series of chromosome segment substitution lines developed between cultivars Nipponbar and Kasalath, one containing a single stem diameter QTL (sdm8; NIL114), and another with four stem diameter QTLs (sdm1, sdm7, sdm8, sdm12; NIL28). Compared with the Nipponbare control, stem diameters were larger in NIL114 and NIL28 by about 7 and 39%, respectively. Pushing resistance in NIL28 was significantly greater than in Nipponbare, but NIL114 was similar to Nipponbare. The two NILs had greater weight of lower stem and culm wall thickness than Nipponbare. NIL28 had higher plant height, which is a negative effect on lodging resistance, than Nipponbare. The non-structural carbohydrate contents of NIL stems were higher than that of Nipponbare, whereas the silicon contents were lower in the NILs, and cellulose contents were lower only in NIL28. The basal internodes of the two NILs were significantly stiffer than those of Nipponbare. These results suggest that increasing stem diameter in rice breeding programs would improve lodging resistance, although the combination of multiple QTLs would be necessary to produce thicker stems with higher pushing resistance, whereas the higher plant height could also result from the combination of multiple QTLs.     

Upgrading the genome of an elite japonica rice variety Kongyu 131 for lodging resistance improvement

Developing a new rice variety requires tremendous efforts and years of input. To improve the defect traits of the excellent varieties becomes more cost and time efficient than breeding a completely new variety. Kongyu 131 is a high-performing japonica variety with early maturity, high yield, wide adaptability and cold resistance, but the poor-lodging resistance hinders the industrial production of Kongyu 131 in the Northeastern China. In this study, we attempted to improve the lodging resistance of Kongyu 131 from perspectives of both gene and trait. On the one hand, by QTL analysis and fine mapping we discovered the candidate gene loci. The following CRISPR/Cas9 and transgenic complementation study confirmed that Sd1 dominated the lodging resistance and favourable allele was mined for precise introduction and improvement. On the other hand, the Sd1 allelic variant was identified in Kongyu 131 by sequence alignment, then introduced another excellent allelic variation by backcrossing. Then, the two new resulting Kongyu 131 went through the field evaluation under different environments, planting densities and nitrogen fertilizer conditions. The results showed that the plant height of upgraded Kongyu 131 was 17%–26% lower than Kongyu 131 without penalty in yield. This study demonstrated a precise and targeted way to update the rice genome and upgrade the elite rice varieties by improving only a few gene defects from the perspective of breeding.     

中国小麦主推品种和育成品系的抗倒伏性评价

倒伏是影响小麦产量的主要因素之一,选育抗倒伏性强的品种是育种研究的重点目标。本研究以528份我国主推的小麦品种和育成品系为材料,对影响小麦倒伏的主要性状进行测定,利用无偏线性估计值进行遗传变异分析、相关分析、主成分分析、线性回归分析和聚类分析,综合评价不同小麦品种的抗倒伏性。研究表明,6个抗倒伏性状在品种间都存在着广泛的遗传变异。基部节间长度、株高与抗推力呈极显著负相关,基部节间直径与抗推力呈极显著正相关,节间长度与节间直径相关性不显著。主成分分析表明,所有性状的信息可以用3个主成分代表,其累计贡献率达83.837%。回归分析结果表明,第1节间直径、第1节间长度、第2节间直径、第2节间长度和株高等5个性状均对抗推力有显著影响。依据抗推力将528份材料聚类为4类,其中第Ⅰ类群有66份小麦品种(系),其抗倒伏能力最强。研究结果为小麦抗倒伏品种利用、抗倒伏育种亲本的筛选以及育种后代品系的选育评价提供理论依据和技术支撑。     

The Use of Mixture Models to Detect Effects of Major Genes on Quantitative Characters in a Plant Breeding Experiment

An analysis based on Elston's model of mixed major locus and polygenic inheritance is extended to include populations of progeny testing such as F(3), B(1s) and B(2s) families derived from F(2) and backcrosses in a cross between two inbred lines. Genetic hypotheses that can be validly tested by the likelihood ratio method in the analysis of a breeding experiment include homogeneity of variances due to environment and/or polygenes with transformable scale effect by Box-Cox power function, random and independent segregation of major genes, invariance of the effects of major genes with population types and additive and dominant models for polygenes. Testing hypotheses in the order suggested here can lead to a gradual simplification of the models and increases the feasibility of the subsequent analysis, but caution must be paid to the possible bias in parameter estimation and hypotheses tests. The procedure is applied to a set of data on plant height of rice with the effects of dwarf genes in crosses among three varieties. Two recessive dwarf genes are shown to be nonallelic and unlinked. One dwarf gene is shown to reduce plant height about 36-56 cm, and another 52-61 cm. The effect of polygenes, estimated as the standard deviation among possible inbred lines derived from these crosses, is about 11.7 cm. Interactions between the dwarf genes and the polygenic background are found, especially for one of the two genes. Both the polygenic effects and the interactions are much smaller than the effects of the major dwarf genes.