水稻第6染色体短臂谷壳硅含量QTL的分解

报道了水稻谷壳硅含量QTLqHUS6的分解研究。针对前期定位的qHUS-6区域,应用在第6染色体短臂RM587-RM19784区间分离、背景基本纯合的F2:3群体,将qHUS-6分解为2个QTL;其中,qHUS-6-1位于目标区间的上部,qHUS-6-2位于下部。同时,筛选出在目标区间内携带更小杂合片段的3个单株。其中,2株覆盖qHUS-6-1区间,各自交产生了1个F2:3群体,应用这2个群体将qHUS-6-1定位于RM510和RM19417之间约147.0kb的区间内;另1株覆盖qHUS-6-2区间,自交产生了1个F2:3群体,从中挑选出在qHUS-6-2区间具有不同基因型组成的5套F3株系,将qHUS-6-2分解为qHUS-6-2a和qHUS-6-2b,分别位于RM19706-RM19795和RM314-RM19665区间。     

测定水稻硅含量的一种简易方法

 介绍了一种简易的测量水稻硅含量的方法，其主要特点为准确、方便、能批量化测定水稻硅含量。以珍汕97B/密阳46重组自交系群体，两次重复，共498个株系的谷壳、剑叶和茎秆为材料，验证了该方法的可靠性。每份材料测定2份样品，两次测定结果达极显著正相关（P<0.01），线性相关系数分别为0.97954、0.97026和0.98848。取10份代表性样品进行了高温碱熔解法的测定，两种方法测定结果高度一致， r = 0.9993。     

Rapid Determination of Silicon Content in Rice

A method for rapid determination of silicon content in rice was introduced. The reliability of this method was verified by using a recombinant inbred line （RIL） population of rice cross Zhenshan 97B / Milyang 46. Two hundred and forty-nine RILs were transplanted in two replications. Simple correlation coefficients on the silicon content in the hull, flag leaf and stern in rice between duplicate samples of 498 rice materials were 0.97954, 0.97026 and 0.98848, respectively. Ten representative samples were selected for measurement using the high-temperature alkaline fusion method. Simple correlation coefficient between the silicon contents determined by the high-temperature alkaline fusion method and by the present method is 0.9993.     

水稻第6染色体短臂上株高QTL qPH6-1的精细定位

 应用衍生于水稻剩余杂合体的分离群体,开展株高QTL的检测和精细定位。应用1个在水稻第6染色体短臂约7.3 Mb区间分离、背景基本纯合的F2:3群体,种植于海南、浙江两地,检测到2个控制株高的QTL;然后,针对两地间作用稳定的qPH6 1,筛选出3个杂合区间缩小且呈阶梯状排列的单株,衍生F2群体,进一步验证了qPH6 1的作用,并将其界定于距离为96.4 kb的SSR标记RM3414和RM19417之间;最后,应用分离区间进一步缩小且呈阶梯状排列的3个F2群体,将qPH6 1定位于距离为51.7 kb的STS标记Si2925和SSR标记RM19417之间。基因组位置比较结果显示,该基因与所有已定位或克隆的水稻矮秆、半矮秆基因均非等位。     

不同生育时期施用硅肥对优质食味粳稻产量和品质的影响

探究不同生育时期施用硅肥对优质食味粳稻产量和品质的影响,可为优质食味粳稻合理施硅提供理论基础和技术支撑。以优质食味粳稻品种南粳9108为试验材料,以不施肥（CK1）和常规施肥（CK2）为对照,在常规施肥的基础上分别设置硅肥浸种（T1）、硅肥浸种+苗期喷施硅肥15 kg/hm²（T2）、硅肥浸种+拔节期喷施硅肥15 kg/hm²（T3）、硅肥浸种+抽穗期喷施硅肥15 kg/hm²（T4）、硅肥浸种+苗期20%、拔节期30%、抽穗期50%喷施硅肥15 kg/hm²（T5）、硅肥浸种+苗期1/3、拔节期1/3、抽穗期1/3喷施硅肥45 kg/hm²（T6）处理,比较不同施硅方式对水稻产量和品质的影响。结果表明,T6处理产量最高,其次是T5、T4、T3处理,T1和T2处理产量较CK2增幅较小;不同硅肥处理单位面积穗数、每穗颖花数、千粒重和结实率较不施硅处理均有不同程度提高。施硅对产量构成因素的影响大小依次为每穗颖花数>单位面积穗数>千粒重>结实率,T3、T5和T6处理的单位面积穗数和每穗颖花数增幅较大。与CK2相比,T3、T4、T5和T6处理增加了水稻库容量、源强度及源库比,提高了糙米率、精米率和整精米率,同时降低了垩白粒率、垩白度及蛋白质含量。T6处理稻米的峰值黏度、热浆黏度、崩解值、最终黏度较CK2显著提高,而其他处理对稻米淀粉黏滞谱特性的影响不显著。主成分分析结果表明,不同硅肥处理对水稻产量和品质影响的大小依次为T6>T5>T4>T3>T2>T1。综合考虑硅肥投入、水稻产量及品质,在常规施肥加硅肥浸种基础上,优质食味粳稻推荐硅肥施用方式为：喷施量15 kg/hm²,苗期占20%、拔节期占30%、抽穗期占50%。     

A Point Mutation in an F-Box Domain-Containing Protein Is Responsible for Brown Hull Phenotype in Rice

The accumulation of pigments affects the color of rice hulls while only limited information is known about its underlying mechanisms. In the present study, a rice brown hull 6(bh6) mutant was isolated from an ethane methyl sulfonate(EMS)-induced IR64 mutant bank. Brown pigments started to accumulate in bh6 rice hulls after heading and reached a higher level in mature seeds. Some major agronomic traits including panicle length and 1000-grain weight in bh6 were significantly lower than those in its corresponding wild type IR64, while other agronomic traits such as plant height, growth duration and seed-setting rate were largely similar between the two genotypes. The analysis of pigment content showed that the contents of total flavonoids and anthocyanin in bh6 hulls were significantly higher than those in IR64 hulls. Our results showed that the brown hull phenotype in bh6 was controlled by a single recessive gene which locates on the long arm of chromosome 9. Sequencing analysis detected a single base substitution(G/A) at position 1013 of the candidate gene(LOC_Os09g12150) encoding an F-box domain-containing protein(FBX310). Functional complementation experiment using the wild type allele can rescue the phenotype in bh6. Thus, we named this mutated gene as Os FBX310~(bh6), an allele of OsFBX310 functioning as an inhibitor of brown hull. The isolation of Os FBX310~(bh6) and its wild type allele can provide useful experimental materials and will facilitate the studies on revealing the mechanisms of flavonoid metabolism in monocot plants.     

硅肥对北方粳稻产量和品质的影响

 以沈农265和丰优2000为材料,在盆栽条件下,研究了硅肥不同施用量对水稻产量和品质的影响。结果表明,施用硅肥能提高水稻产量,其中有效硅240 kg/hm2使沈农265显著增产,有效硅180 kg/hm2对丰优2000增产效果达到显著水平。从产量构成因素上看,施用硅肥能提高每穴穗数、一次枝梗上籽粒千粒重、二次枝梗上籽粒千粒重、每穗粒数。施用硅肥能显著改善稻米品质,主要表现为稻米的糙米率、精米率和整精米率显著提高,稻米的垩白粒率和垩白度显著降低。相关分析表明,两品种的精米率和整精米率与有效硅施用量呈正相关。沈农265的稻米脂肪酸含量与有效硅施用量呈正相关,食味值与有效硅施用量呈负相关。有效硅施量在0~240 kg/hm2内,两品种直链淀粉含量和蛋白质含量与有效硅施用量相关不显著。综合考虑各处理稻米产量和品质,以180~240 kg/hm2的有效硅施用量较好。     

水稻色素原基因C的精细定位

应用籼稻组合珍汕97B/密阳46的衍生材料,针对水稻第6染色体短臂色素原基因C的可能位置,筛选到在C基因周围区间呈不同基因型组合的7个剩余杂合体,收获种子建立F2∶3群体。在各个植株上,稃尖颜色和叶鞘颜色的表现完全相同。通过各个群体颜色表现与原剩余杂合体基因型的比较,将C基因定位于微卫星标记RM314与RM253之间。在该基础上,应用两个分离群体共1279个样本,经标记检测和连锁分析,进一步将C基因定位于RM111和RM253之间, 与RM111和RM253的遗传距离分别为0.7 cM和0.4 cM。最后,应用区间内的另外6个微卫星标记和1个源于C基因候选基因OsC1的标记,检测在RM111 C基因 RM253区间内发生了重组的22个个体,将C基因定位于一个大小为59.3 kb、涵盖C基因候选基因OsC1座位的区间中。     

Fine Mapping of C(Chromogen for Anthocyanin)Gene in Rice

Seven residual heterozygous lines(RHLs)displaying different genotypic compositions in the genomic region covering probable locations of C (Chromogen for anthocyanin)gene on the short arm of rice chromosome 6 were selected from the progenies of the indica cross Zhenshan 97B/Milyang 46.Seeds were harvested from each of the seven plants,and the resultant F2:3 populations were used for fine mapping of C gene.It was shown in the populations that the apiculus coloration matched to basal leaf sheath coloration in each plant.By relating the coloration performances of the populations with the genotypic compositions of the RHLS,the C locus was located between rice SSR markers RM314 and RM253.By using a total of 1279 F2:3 individuals from two populations showing coloration segregation.the C locus was then located between RM111 and RM253,with genetic distances of 0.7 cM to RM111 and 0.4 cM to RM253.Twenty-two recombinants found in the two populations were assayed with seven more markers located between RM111 and RM253,including six SSR markers and one marker for the C gene candidate,OsC1.The C locus Was delimited to a 59.3-kb region in which OsC1 was located.     

供锌水平对水稻生长和锌积累和分配的影响

通过对3个不同水稻品种（碧玉早糯、26715和浙农921）5个供锌水平（0.0、0.5、2.0、8.0、32.0 μmol/L ZnSO[sub]4[/sub]）处理的水培试验,研究供锌水平对水稻生长和锌积累的影响。结果表明,水稻地上部锌含量随供锌水平的提高而提高,不同的供试品种在不同的生育期地上部锌含量的变化趋势存在差别。不同的供试品种之间籽粒锌的积累差异极显著,精米中的锌含量随供锌水平的提高而提高;在8.0、32.0 μmol/L Zn水平下,籽粒锌含量差异不大,颖壳锌含量则随供锌水平的提高而提高;在≤8.0 μmol/L时,精米锌含量比颖壳高,而在高锌水平（32.0 μmol/L）下,颖壳的锌含量比精米高。     

Distribution Characterization of Leaf and Hull Pubescences and Genetic Analysis of Their Numbers in japonica Rice (Oryza sativa)

Distributions of pubescences on leaf blade and hull in japonica rice were observed under an optical microscope. Numbers of leaf and hull pubescences in P1, P2, F1, B1, B2 and F2 generations were investigated in three combinations of japonica rice (Sidao 10A/Wuyujing 3R, Wuyujing 3A/Sidao 10R and Liuyan 189A/HR-122), and genetic analysis for these two traits were conducted by using the joint analysis method of P1, P2, F1, B2, B2 and F2 generations with the mixed major gene plus polygene inheritance models. Leaf pubescences characterized by swollen base and fine tip distributed regularly on the boundary between dark green stripe and light green stripe of leaf blade. Hull pubescences with various lengths distributed irregularly on the whole hull. Numbers of leaf pubescences in the reciprocal combinations of Sidao 10A/Wuyujing 3R and Wuyujing 3A/Sidao 10R and numbers of hull pubescences in all the three combinations were controlled by one pair of additive major genes plus additive-dominant polygenes. In the combination of Liuyan 189A/HR-122, number of leaf pubescences was controlled by one pair of additive-dominant major genes plus additive-dominant polygenes. Both numbers of leaf and hull pubescences were mainly governed by major genes.     

水稻籽粒钾和蛋白质含量的基因型差异

采用对低钾胁迫敏感程度不同的3种水稻基因型进行田间试验, 以研究水稻籽粒中钾和蛋白质含量。结果表明,低钾胁迫降低水稻籽粒中钾含量和粗蛋白含量,谷壳中钾含量高于米粒中钾含量,米粒中钾含量与粗蛋白含量呈显著正相关。钾高效基因型在低钾胁迫下谷壳和米粒中具有较高的钾相对含量,其米粒中的粗蛋白相对含量也较高。     

硅肥对膜下滴灌水稻生长发育及产量的影响

为确定适合膜下滴灌水稻生长的施硅量,以T-43为材料,研究了不同用量硅肥对膜下滴灌水稻生长发育和产量的影响。结果表明,与对照(硅肥用量为0 kg/hm~2)相比,施硅有利于水稻分蘖早发快发,减少无效分蘖,提高水稻生育后期叶片叶绿素含量;施硅还可以增加水稻秆型指数,增强植株抗倒伏能力,135 kg/hm~2的施硅量秆型指数最大;90 kg/hm~2的施硅量使水稻后期冠层有效辐射吸收量降低变慢,有利于水稻抽穗后更好的利用光能进行有机物生产;硅肥主要是通过影响穗长、实粒数和结实率来促进产量的提高,施硅量135 kg/hm~2的处理产量最高,但与施用量为90 kg/hm~2的处理差异不显著。综上所述,施用硅肥有利于膜下滴灌水稻的生长,但从经济角度考虑,以施用量为90 kg/hm~2较为适宜。     

水稻耐金属离子胁迫的QTL分析

【目的】 本研究旨在筛选与水稻苗期耐不同金属离子连锁的分子遗传标记,为探讨水稻耐不同金属离子胁迫的遗传研究提供参考。【方法】 以典型籼粳交(春江06/台中本地1号)双单倍体(DH)群体为材料,系统考查该群体及其双亲耐4种金属离子(Fe²⁺、Cd²⁺、Al³⁺、Na⁺)胁迫的情况,利用业已构建并完善的该群体加密的分子连锁图谱,对耐这4种金属离子胁迫的QTL进行检测分析。另外,利用实时定量PCR技术检测处理前后相关基因的表达变化情况。【结果】 发现耐各种金属离子胁迫的QTL共8个,分别位于水稻第1、2、4、6、9、10和11染色体上,其中Fe²⁺处理后检测到的QTL贡献率最大,达到24.47%(阈值为7.78),位于第1染色体上RM1297–RM1061,同时对该区间与耐胁迫相关基因的表达分析发现这些基因在处理前和处理后表达水平存在不同程度的差异;Cd²⁺处理后检测到1个QTL,位于第1染色体上;Al³⁺处理后检测到QTL共5个,分别位于第2、4、6、10、11染色体上;Na⁺处理后检测到QTL有1个,位于第9染色体上。【结论】 根据不同金属离子胁迫处理后DH群体的表型差异进行QTL分析,发现耐各种金属离子胁迫的QTL共8个,并初步定位于各染色体的遗传标记区间,这为精细定位并克隆相应QTL,进而探明水稻耐金属离子胁迫QTL的分子调控机制奠定了基础。     

QTL mapping for crude protein and protein fraction contents in rice (Oryza sativa L.)

Improvement of rice storage proteins is important in rice breeding for high nutritional quality. Seventy-one recombinant inbred lines (RIL) derived from a cross between japonica variety Asominori and indica variety IR24 were used to study the inheritance of crude protein and protein fraction contents in rice. A total of 16 QTL were identified and mapped on eight chromosomes. Several QTL affecting contents of different protein fractions were mapped in the same chromosomal region. In particular, two QTL with a significant contribution were identified to simultaneously affect prolamin and glutelin contents. One QTL denoted as qCP-12 affecting crude protein content (CP) was located in the same region as QTL qGLT-12 affecting glutelin content, in agreement with the positive correlation between glutelin level and protein content. QTL with larger genetic effects were further confirmed using two sets of chromosome segment substitution lines (CSSL), where Asominori and IR24 were used as the recurrent parents. By QTL comparative analysis, two QTL for CP, three for globulin content and one for prolamin content were located in the vicinity of CP QTL previously identified in polished rice. Application of these results in rice breeding is discussed.     

Fine Mapping of C（Chromogen for Anthocyanin） Gene in Rice

Seven residual heterozygous lines (RHLs) displaying different genotypic compositions in the genomic region covering probable locations of C (Chromogen for anthocyanin) gene on the short arm of rice chromosome 6 were selected from the progenies of the indica cross Zhenshan 97B/Milyang 46. Seeds were harvested from each of the seven plants, and the resultant F_2:3 populations were used for fine mapping of C gene. It was shown in the populations that the apiculus coloration matched to basal leaf sheath coloration in each plant. By relating the coloration performances of the populations with the genotypic compositions of the RHLs, the C locus was located between rice SSR markers RM314 and RM253. By using a total of 1279 F_2:3 individuals from two populations showing coloration segregation, the C locus was then located between RM111 and RM253, with genetic distances of 0.7 cM to RM111 and 0.4 cM to RM253. Twenty-two recombinants found in the two populations were assayed with seven more markers located between RM111 and RM253, including six SSR markers and one marker for the C gene candidate, OsC1. The C locus was delimited to a 59.3-kb region in which OsC1 was located.     

两个垩白突变体的鉴定及突变基因的图位克隆

【目的】研究两个水稻垩白突变体胚乳垩白的形成机制,为稻米品质改良提供理论基础。【方法】以从中花11的EMS突变体库中筛选出的两个稳定遗传的垩白突变体eb6、eb7为材料,对其进行农艺性状、稻米理化性质和遗传学分析,并利用eb6与南京11衍生的F2群体对控制垩白的基因进行图位克隆。同时对候选基因的表达模式及淀粉合成相关基因在突变体及野生型中表达情况进行了分析。【结果】与野生型相比,两个突变体胚乳中央部位呈现白色且不透明,淀粉复合颗粒形状不规则且排列疏松,而突变体胚乳边缘部位与野生型无异,都为半透明状,淀粉复合颗粒呈多面体且排列致密。相对于野生型,突变体eb6、eb7成熟种子中的直链淀粉含量和胶稠度显著降低,蛋白质含量显著升高。RVA谱分析显示突变体淀粉黏滞性明显低于野生型。同时,支链淀粉聚合度分析显示突变体eb6中聚合度(DP)小于16的短链显著增加,DP为16~23的中长链显著减少。遗传分析表明突变体eb6、eb7胚乳垩白表型由单隐性核基因控制,并且它们为一对等位突变体。利用突变体eb6与南京11杂交衍生的F2群体将突变体基因定位在第1染色体长臂上物理距离86.6 kb的区间内。该区间包含22个开放阅读框(ORF),其中ORF13编码腺苷二磷酸葡萄糖焦磷酸化酶大亚基2(Os AGPL2)。序列分析发现eb6与eb7分别在Os AGPL2第3、第7外显子上发生1个单碱基替换,并分别导致一个氨基酸替换。RT-PCR及原位杂交结果显示,Os AGPL2主要在水稻发育中的籽粒中表达。同时Os AGPL2的突变导致了多个淀粉合成相关基因在水稻籽粒灌浆过程中的表达模式发生改变。【结论】突变体eb6、eb7籽粒胚乳出现严重垩白表型为Os AGPL2突变所致。本研究进一步证明了Os AGPL2在调控水稻籽粒胚乳中淀粉的合成、淀粉复合颗粒的形成及稻米理化性质的平衡中起着重要的作用。     

粳稻叶片茸毛和谷壳稃毛的分布特征及毛数的遗传分析

在光学显微镜下观察了粳稻叶片茸毛在叶面上的分布特征和谷壳稃毛在谷壳上的分布特征。调查了粳稻3个杂交组合的P1、P2、F1、B1（F1/P1)、B2(F1/P2) 和F2共6个世代的叶片茸毛数和谷壳稃毛数,运用主基因+多基因混合遗传模型6个世代联合分析的方法,对这两个性状进行了遗传分析。结果发现,叶片茸毛有规律地分布在叶面的暗绿条带和淡绿条带的交界线上,形状是基部膨大,顶端细小;谷壳稃毛无规律地分布在整个谷壳上,长短不一。在泗稻10号A/武育粳3号R与武育粳3号A/泗稻10号R正反交组合中叶片茸毛数均受1对加性主基因+加性 显性多基因共同控制,在六盐189A/HR 122组合中叶片茸毛数受1对加性 显性主基因+加性 显性多基因共同控制; 在3个杂交组合中谷壳稃毛数均受1对加性主基因+加性 显性多基因共同控制;叶片茸毛数和谷壳稃毛数都以主基因遗传为主。     

广东雷州杂草稻黑壳基因Bh4和红皮基因Rc的基因型鉴定

为测定雷州杂草稻的黑色颖壳基因Bh4和红色果皮基因Rc的基因型,本研究对雷州10个杂草稻群体的100个单株的种子的颖壳和果皮颜色进行表型观察,同时通过PCR扩增及序列分析对Bh4和Rc的基因型进行鉴定。结果表明,雷州杂草稻中黑色颖壳和黄色颖壳的种子几乎各占一半,果皮颜色则以红色为主,有少量白色和浅红色果皮的种子。雷州杂草稻的Bh4和Rc的基因型与表型鉴定结果一致。57份黑色颖壳杂草稻的Bh4基因型为39个野生型和18个杂合型,25份黄色颖壳和18份黄色带黑斑颖壳杂草稻的Bh4基因型均为缺失22 bp的突变型;90份红色果皮杂草稻的Rc基因型为73个野生型和17个杂合型,4份白色和6份浅红色果皮杂草稻的Rc基因型均为缺失14 bp的突变型。本研究为分析雷州杂草稻的Bh4和Rc基因的来源提供了分子基础。     

水稻第6染色体短臂株高及产量性状QTL的分解

针对第6染色体短臂上一个对产量性状遗传具有重要作用的区间RM587-RM19715,从珍汕97B/密阳46重组自交系群体中筛选到1个剩余杂合体,自交衍生获得一个由195个个体组成的F2群体,检测控制株高和产量性状的QTL。经分析,在目标区间的上部和下部分别检测到1个QTL簇,分别对除单株穗数以外的产量性状因子具显著作用,单个QTL对群体性状表型变异的贡献率为5.0%~55.5%。将第6染色体上的产量性状QTL分解到更小的区间中,为产量性状QTL的精细定位和克隆打下了基础。