水稻糯性突变对淀粉理化特性的影响

比较了籼、粳两个糯性突变品种与原亲本品种淀粉的理化特性。糯性突变后除了直链淀粉含量显著降低外,淀粉的糊化温度变高、胶稠度增大、RVA谱(rapid viscosity analyzer)参数下降。RVA谱参数降低的幅度从高到低依次是消减值>回复值>冷胶粘度>热浆粘度>最高粘度>崩解值,表明控制直链淀粉含量的Wx位点对淀粉糊化特性和粘滞性是有影响的,存在多效性。同时,两个糯稻品种之间的RVA谱差异较大,其中崩解值和最高粘度差值均大于糯性突变对它们的影响,说明除直链淀粉外,支链淀粉也影响淀粉的理化特性。对Wx位点鉴定的结果显示扬辐糯4号wx基因第一内含子剪切位点＋1位的碱基由原品种的G突变成了T,扬糯2号与其原品种相同(均为T);两个糯稻突变品种wx基因前导区的(CT)n微卫星重复序列与原品种相比都发生了变化。     

分子标记辅助选择与花药培养相结合快速聚合水稻白叶枯病抗性基因

以受微效多基因控制白叶枯病抗性的五丰占2号和主基因[i]xa5[/i]控制的IRBB5为材料,应用分子标记辅助选择、花药培养和回交技术研究了主基因与微效多基因聚合的途径。经查阅GeneBank中RG556的序列,利用在线引物设计软件Primer 3设计了一条新的R引物：5′CCAGACACCACTGCACATTC3′,克服了原引物Tm值相差太大,扩增效率低的缺点,使PCR扩增效率提高了近10倍。采用病斑长度与五丰占2号相近且携有xa5基因的五丰占2号[sup]2[/sup]/IRBB5 B[sub]1[/sub]F[sub]1[/sub]植株进行花药培养,获得30株二倍体植株,其中16株携有[i]xa5[/i]基因。携有[i]xa5[/i]基因的植株的平均病斑长度仅0.1 cm,对白叶枯病的抗性明显强于双亲。成功地聚合了白叶枯病抗性主基因和微效多基因。从配组到获得主基因和微效多基因聚合材料仅用两年时间,比常规育种方法缩短1~2年。     

转抗除草剂基因水稻与亲本的稻米理化特性及主要营养成分比较

对转抗除草剂基因水稻与原亲本稻米的理化特性和关键性营养成分进行了比较,发现在最高粘度、热浆粘度和冷胶粘度等RVA谱特征值上两者有显著差异,但在表观直链淀粉含量、胶稠度和淀粉粘滞特性的崩解值和消减值等淀粉食用品质方面基本相同;在粗蛋白、粗脂肪、总灰分、氨基酸和矿物质等关键性营养成分上不存在显著差异。表明在水稻遗传转化操作过程中,T-DNA插入并未显著改变原亲本的上述品质性状。     

太湖流域粳稻地方品种韭菜青的苗期耐盐性遗传分析

为了研究太湖流域强耐盐粳稻地方品种韭菜青的苗期耐盐性遗传机理,配制了韭菜青/IR26籼粳杂交组合,在盐胁迫条件下,运用剥蘖分株法鉴定了P[sub]1[/sub]、P[sub]2[/sub]、F[sub]1[/sub]和F[sub]2[/sub]苗期的盐害级别和根系Na+/K+两个耐盐相关指标。结果表明,在0.8%的NaCl胁迫下,亲本的两个耐盐相关指标有明显的差异,F[sub]1[/sub]均表现出超亲优势,F[sub]2[/sub]呈正态混合分布。采用主基因 多基因遗传模型进行遗传分析,发现韭菜青苗期的盐害级别和根系Na[sup]+[/sup]/K[sup]+[/sup]均由2对主效基因控制,并存在多基因修饰。盐害级别的主基因遗传率仅为16.10%,而根系Na[sup]+[/sup]/K[sup]+[/sup]的主基因遗传率高达50.78%,两个指标的主基因加多基因共同解释的遗传率都在70%以上。     

空间诱变水稻矮秆突变体CHA-1对赤霉素的反应及其遗传分析

应用不同浓度的赤霉素（GA[sub]3[/sub]）溶液在苗期处理CHA-1特华占原种、日本晴、矮脚南特等4个材料,研究它们对GA[sub]3[/sub]反应的敏感性。结果表明,经空间诱变的矮秆突变体在苗期对GA[sub]3[/sub]的反应敏感,但与已知带[i]sd-1[/i]基因的籼、粳稻代表品种矮脚南特、日本晴敏感性有差异。进一步的遗传分析表明,控制CHA-1的一对隐性矮秆基因与[i]sd-1[/i]基因不等位。     

水稻孕穗期不同叶位叶片的气体交换与叶绿素荧光特性

研究了6个水稻品种孕穗期不同叶位的气体交换和叶绿素荧光特性,结果表明净光合速率([i]P[/i][sub]n[/sub]）、气孔导度（[i]g[/i]）、蒸腾速率（[i]E[/i]）和水分利用效率（[i]WUE[/i]）均随叶位降低而明显下降;而胞间CO[sub]2[/sub]浓度（[i]C[/i][sub]i[/sub]）基本维持不变,说明叶片衰老时[i]P[/i][sub]n[/sub]的下降主要不是气孔因素引起的。叶绿素含量降低与[i]P[/i][sub]n[/sub]下降的表现一致,叶绿素荧光分析表明[i]F[/i][sub]v[/sub]/[i]F[/i][sub]m[/sub]和[i]F[/i]'[sub]v[/sub]/[i]F[/i]'[sub]m[/sub]没有随叶位发生变化,而PSⅡ的光化学效率[i]Ф[/i][sub]PSⅡ[/sub]和电子传递速率([i]ETR[/i]）随叶位降低明显下降,这表明可能是PSⅠ电子传递受阻导致CO[sub]2[/sub]同化下降。     

江苏省稻瘟病菌群体分布及优势小种的毒力研究

用中国7个统一鉴别品种对2000~2002年江苏省5大稻区采集的342个稻瘟病菌单孢分离菌株进行了小种鉴定。结果表明3年累计出现7群30个小种,其中ZG1为优势小种,3年的频率分别为65.00%、56.90%和60.38%,ZB群和ZC群生理小种也是重要的小种类型,有较高的出现频率。用130个ZG1小种的菌株对日本13个已知抗性基因品种进行毒力测定,出现42种类型毒力,其中30.77％的ZG1小种的菌株同时对品种新2号（Pita-k[sup]s[/sup],Pi-sh）、K1（Pi-ta）、Pi-4号（Pi-ta[sup]2[/sup],Pi-sh）致病,为优势毒力类型, 表明品种新2号（Pita-ks,Pi-sh）、K1（Pi-ta）、Pi-4号（Pi-ta[sup]2[/sup],Pi-sh)与江苏省稻瘟病菌有较高的适合度。品种K3（Pi-k[sup]h[/sup]）在江苏省有较好的抗性,其抗性频率为100%。用两种毒力类型的不同菌株2003-184(ZC[sub]5[/sub])和 2003-14-1(ZG[sub]1[/sub]) 混合接种13个日本已知抗性基因品种后,发现对上述两个单个菌株表现抗病的K3（Pi-k[sup]h[/sup]）品种表现为感病,提示不同毒力的菌株之间的相互作用导致菌株毒力的改变可能是品种抗性丧失的原因之一。     

拮抗细菌Bacillus subtilis B5423-R 抑制水稻纹枯病的阈值群体数量

 在菲律宾国际水稻研究所实验农场田间条件下，通过人工接种Rhizoctonia solani,采用不同的方法(喷雾次数、间隔期)喷雾B5423-R（B5423的标记菌株，为利福平Rifampicin抗性的突变体），在接种1 d后，采用2 d喷1次、连续3～4次喷雾的方法，使B5423 R在稻株上的群体数量在6～7 d内维持在1×10[sup]6[/sup]cfu/g以上，在25 d内显著地降低相对病斑高率；而采用5 d喷1次、连续3～4次喷雾的方法，B5423-R在稻株上的群体数量在15 d内维持在1×10[sup]5[/sup]～106cfu/g，在25 d内相对病斑高率未能显著降低。表明在纹枯病侵染早期的6～7 d内，在稻株上B5423 R维持在1×10[sup]6[/sup]cfu/g以上是抑制纹枯病的阈值群体数量。     

稻米淀粉粘滞性谱特征的表现及其遗传

通过对水稻F[sub]2[/sub]无性系群体及大量品种（系）的RVA谱的测定,研究了稻米淀粉RVA谱的类型及遗传。结果表明：（1）RVA谱可分为A~F六种类型,糯稻的特征图谱为A型,籼稻的特征图谱有B、C、D、E、F五种类型,粳稻有D、E、F三种类型。（2）稻米的表观直链淀粉含量（AAC）不同,RVA谱则不同。稻米的AAC相同,RVA谱也可不同,在低、中AAC类型中,RVA谱的差异能反映品种食味品质的差异。（3）4个杂交稻组合的RVA谱表现出相同的特点,除最高粘度和起浆温度外,其他特征值均介于双亲之间,表明杂交稻米品质改良要求不育系与恢复系的改良同步进行。（4）在F[sub]2[/sub]无性系群体中,RVA谱各特征均呈连续分布,最高粘度是典型的数量性状遗传,热浆粘度、冷胶粘度、崩解值、消减值、回复值都可能是由一对主效基因和若干微效基因共同控制的遗传。     

1-磷酸甘露醇脱氢酶基因转化水稻的研究

摘要：PCR和Southern blotting检测表明,来自大肠杆菌的[i]mtlD[/i]基因已通过农杆菌介导整合进水稻基因组。[i]mtlD[/i]基因在T1代出现分离, T2代出现纯系。在0.75% NaCl胁迫下,7个转基因T3代株系都能检测到mtlD酶活性,与对照相比细胞膜的相对电导率和大分子渗漏值明显降低。部分转基因株系能在 1.0% NaCl浓度下正常生长,而对照在0.5% NaCl浓度下已不能存活。通过有性杂交途径实现了[i]mtlD[/i]和[i]gutD[/i]两个基因的聚合,部分杂交后代株系能在1.25%NaCl胁迫下正常生长结实。     

QTL Mapping for Rice RVA Properties Using High-Throughput Re-sequenced Chromosome Segment Substitution Lines

The rapid visco analyser （RVA） profile is an important factor for evaluation of the cooking and eating quality of rice. To improve rice quality, the identification of new quantitative trait loci （QTLs） for RVA profiling is of great significance. We used a japonica rice cultivar Nipponbare as the recipient and indica rice 9311 as the donor to develop a population containing 38 chromosome segment substitution lines （CSSLs） genotyped by a high-throughput re-sequencing strategy. In this study, the population and the parent lines, which contained similar apparent amylose contents, were used to map the QTLs of RVA properties including peak paste viscosity （PKV）, hot paste viscosity （HPV）, cool paste viscosity （CPV）, breakdown viscosity （BKV）, setback viscosity （SBV）, consistency viscosity （CSV）, peak time （PET） and pasting temperature （PAT）. QTL analysis was carried out using one-way analysis of variance and Dunnett＇s test, and stable QTLs were identified over two years and under two environments. We identified 10 stable QTLs： qPKV2-1, qSBV2-1; qPKV5-1, qHPV5-1, qCPV5-1; qPKV7-1, qHPV7-1, qCPV7-1, qSBV7-1; and qPKV8-1 on chromosomes 2, 5, 7 and 8, respectively, with contributions ranging from -95.6% to 47.1%. Besides, there was pleiotropy in the QTLs on chromosomes 2, 5 and 7.     

杂交稻淀粉RVA谱特征值的配合力及杂种优势研究

以6个籼型三系杂交稻不育系和5个恢复系按不完全双列杂交配制的30个杂交稻组合及其亲本为材料,对其淀粉RVA谱特征值进行了测定和分析。结果表明:淀粉RVA谱特征值在各组合间均存在极显著遗传差异,其加性效应和非加性效应对杂交稻淀粉RVA谱特征值均存在极显著的影响,并以加性效应为主;杂交稻亲本RVA谱特征值一般配合力(GCA)效应高的,其后代特殊配合力(SCA)效应不一定高;杂交稻母本RVA谱特征值GCA方差在总的基因型方差中所占比重远远大于父本和组合SCA方差,表明杂交稻淀粉RVA谱特征值主要受不育系的影响;杂交稻淀粉RVA谱特征值的竞争优势与母本的GCA极显著正相关,且决定系数较高,表明杂交稻母本的一般配合力高,其所配杂交稻组合的竞争优势也强;杂交稻淀粉RVA谱特征值的峰值黏度存在明显的超高亲优势;糊化开始温度存在超低亲优势;其他性状存在中亲优势。     

稻米淀粉黏滞性(RVA谱)特征值的遗传及QTL定位分析

利用武育粳3号/Aichi 106构建的F2 群体,研究了精米粉RVA谱8个特征值: 最高黏度(peak viscosity,PKV)、热浆黏度(hot paste viscosity,HPV)、冷胶黏度(cool paste viscosity,CPV)、崩解值(breakdown viscosity,BDV)、消减值(setback viscosity,SBV)和回复值(consistence viscosity,CSV)、峰值时间（peak time, PeT）、糊化温度（pasting temperature, PaT）的遗传规律。结果表明, PKV为由微效基因控制的数量性状;HPV、CPV、SBV、BDV、CSV可能是由一对主基因和微效多基因共同控制的数量性状;PeT可能由两对主基因控制,并且还受微效多基因的影响。用热研2号/密阳23构建重组自交系群体,于2005-2006年对稻米淀粉RVA谱8个特征值进行了数量性状基因座QTL定位和分析,两年试验共检测到34个QTL,分布在第1、2、3、4、6、7、8共7条染色体上;两年同时检测到的QTL共8个,其中qHPV6、 qCPV6、 qCSV6、 qSBV6、qBDV6位于第6染色体的Wx基因附近;qHPV2、qCSV2位于第2染色体RM341与RM475标记之间,qCPV2位于RM573与RM250标记之间。第2染色体上与支链淀粉合成有关的两个基因与RVA谱特征值有密切关系。     

Inheritance Analysis and QTL Mapping of Rice Starch Viscosity (Rapid Visco Analyzer Profile) Characteristics

The rice starch viscosity characteristics, which can be indicated by Rapid Visco Analyzer profile (RVA profile), have been proved useful for the evaluation of cooking and eating quality in rice breeding program. To study the inheritance of the RVA profile, an F2 population of Wuyujing 3/Aichi 106 was used. The results indicated that the peak viscosity (PKV) was a typical quantitative character, and the hot paste viscosity (HPV), cool paste viscosity (CPV), setback viscosity (SBV), breakdown viscosity (BDV), and consistence viscosity (CSV) might be controlled by one major gene and several minor genes.To elucidate the genetic basis of the paste viscosity characteristics, a recombinant inbred line (RIL) population derived from Nikken 2/Milyang 23 and its genetic linkage map were used to map the QTLs controlling RVA profiles in 2005 and 2006. A total of 34 QTLs distributed on chromosomes 1, 2, 3, 4, 6, 7 and 8 were detected, including 19 and 15 QTLs in 2005 and 2006,respectively. Eight QTLs were both detected in the two years, qHPV6, qCPV6, qCSV6, qSBV6, and qBDV6were located on chromosome 6, while qHPV2, qCSV2, and qCPV2 were on chromosome 2.     

氮肥追施量对香稻1601品质和淀粉RVA谱特征值的影响

以自育品系香稻1601为材料,研究不同氮肥追施量对其品质和淀粉RVA谱特征值的影响,为探明氮肥施用量与香稻品质关系提供数据支撑。试验结果表明:随着氮肥追施量的增加,稻米蛋白质含量增加,食味品质下降趋势,淀粉RVA谱曲线整体下降,峰值黏度、热浆黏度、冷胶黏度均呈下降趋势,崩解值和消减值先升高后降低,糊化温度和峰值时间无显著差异。相关分析表明,峰值黏度和冷胶黏度与大米蛋白质含量极显著负相关,相关系数-0.889和-0.843,与食味值存在极显著或显著正相关,相关系数0.896和0.763。     

Starch digestibility and the estimated glycemic score of different types of rice differing in amylose contents

Three types of rice cultivars (indica, japonica and hybrid rice) with four levels of amylose were selected for assessing variability in starch digestibility. A vitro enzymatic starch digestion method was applied to estimate the glycemic index in vivo based on the kinetics of starch hydrolysis in vitro. The results indicated that significant differences in term of glycemic response were observed in three types of rice. Amylose content had an obviously impact on the estimated glycemic score (EGS) value and resistant starch (RS) content. The contents of RS were increased with the increasing amylose in the same type of rice. Japonica rice was significantly lower in RS content compared to indica rice and hybrid rice with similar amylose. The high amylose rice cultivar ZF201, which was characterized by low major RVA parameters, i.e. peak viscosity (PKV), hot paste viscosity (HPV) and cool paste viscosity (CPV), were obviously higher in RS content and lower in EGS. The retrogradation of cooked rice led to a reduction of HI and EGS of all varieties. Starch hydrolysis tends to be more quick and complete for the waxy and low amylose rice than for the intermediate and high amylose rice.     

优质水稻品种越光(Koshihikari)中控制稻米淀粉RVA谱特征值的QTL分析

利用南京11×Koshihikari RIL群体在南京两年的种植结果,检测与稻米淀粉RVA谱特征值峰值黏度、最低黏度、最终黏度、崩解值、消减值、峰值时间和糊化温度相关加性和上位性效应QTL,并分析其表达稳定性。结果表明,7个稻米淀粉RVA谱特征值在两年中均呈连续分布,且存在超亲遗传现象。共检测到8个具有加性效应的QTL与稻米6项淀粉RVA谱特征值有关,两年中能重复出现的QTL有6个,即qTPV-6、qFPV-6、qBDV-6、qSBV-6、qPKT-6和qPT-6,分别控制最低黏度、最终黏度、崩解值、消减值、峰值时间和糊化温度,两年的平均贡献率分别为46.4%、60.3%、31.1%、71.9%、38.5%和12.4%,而qSBV-8和qPT-5仅在1年中被检测到,环境稳定性差。此外,还检测到5对影响稻米淀粉RVA谱特征值的QTL具有上位性效应,互作既发生在相同染色体上也发生在不同染色体之间,但贡献率较小,且受环境的影响较小。控制稻米淀粉RVA谱特征值的稳定、主效QTL,可为Koshihikari中稻米优良蒸煮食味品质相关基因的育种利用提供基础。     

Allelic Effects on Eating and Cooking Quality of SSⅡa and PUL Genes Under Wxmp Background in Rice

【Objective】Our aim is to analyze the genetic effects of soluble starch synthase gene SSⅡa and debranching enzyme gene PUL on eating and cooking quality under the same main gene background of Wxmp, so as to lay a theoretical basis for rice quality improvement. 【Method】In this study, a semi-glutinous rice line Ning 0145 and japonica rice variety Wuyunjing 21 were crossed to obtain F2 population and F3 lines. There was polymorphism in soluble starch synthase gene SSⅡa and debranching enzyme gene PUL but no polymorphism in other starch synthase related genes between the two parents. With molecular markers, some F2 plants and F3 lines containing Wxmp gene were selected and divided into four genotypes, SSⅡanPULn, SSⅡanPULw, SSⅡawPULnand SSⅡawPULw(n and w indicated that the genes were contributed by Ning 0145 and Wuyunjing 21, respectively). The allelic effects of SSⅡa and PUL genes on eating and cooking quality under the same Wxmp gene background was investigated by analyzing the eating and cooking quality and its differences among different genotypes. 【Result】There were significant differences for eating and cooking quality among genotypes of different parental origins. The allelic gene SSⅡaw and PULw from Wuyunjing 21 increased amylose content by 0.29%–1.00% and 0.62%–1.18% respectively, and the effect of PUL was greater than that of SSⅡa. There was interaction between SSⅡa and PUL genes. The SSⅡaw and PULw genes also decreased gel consistency and breakdown viscosity, increased hot paste viscosity, cool paste viscosity, setback viscosity and consistency viscosity, but had little effect on gelatinization temperature, peak viscosity and peak time. 【Conclusion】The genetic effects of SSⅡa and PUL genes on cooking and eating quality of rice under the background of Wxmp gene were clarified. The results lay a theoretical basis for improving rice quality by molecular marker-assisted selection of SSⅡa and PUL genes.