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.     

一个水稻颖壳扭曲突变体的遗传分析与基因定位

 从水稻育种后代材料中获得1个颖壳扭曲突变体Osth (twisted hull)。遗传分析结果表明,该突变性状由单核基因隐性突变造成。以突变体与颖壳正常籼稻R725杂交的F2群体为基因定位群体,利用SSR标记将突变位点定位在第2染色体上的SSR标记RM14128与RM208之间,遗传距离分别为1.4 cM 和2.7 cM。这些结果为该基因的精细定位和克隆以及研究水稻花发育的分子机理奠定了基础。     

水稻斑点叶突变体hm197的鉴定及其基因定位

通过双环氧丁烷 (diepoxybutane)诱变籼稻品种IR64获得遗传稳定的水稻褐色斑点叶突变体hm197。在自然条件下,该突变体褐色斑点自播种后10周开始于叶尖出现,而后慢慢扩散至全叶。遗传分析表明,该褐色斑点性状受一对隐性核基因控制,暂名spl^hm197,并将其定位在第4染色体长臂上140 kb的区段内。与野生型IR64相比,突变体株高、结实率和千粒重等农艺性状均显著下降。遮光处理表明,hm197褐色斑点的形成受自然光照的诱导。此外,hm197光合色素含量和光合效率也比野生型显著降低。组织化学分析表明,突变体中有过氧化氢和大量超氧阴离子O_{2 ?}的沉积。与IR64相比,hm197叶片中清除氧自由基酶系统中SOD和APX活性极显著上升,其余均极显著下降,同时伴随总可溶蛋白含量下降以及MDA含量上升,hm197表现出早衰迹象。抗病性鉴定表明,与野生型相比突变体对白叶枯病菌的抗性显著增强。     

Genetic Analysis and Gene Mapping of Light Brown Spotted Leaf Mutant in Rice

A light brown spotted-leaf mutant of rice was isolated from an ethane methyl sulfonate (EMS)- induced IR64 mutant bank. The mutant, designated as lbsl1 (light brown spotted-leaf 1), displayed light brown spot in the whole growth period from the first leaf to the flag leaf under natural summer field conditions. Agronomic traits including plant height, growth duration, number of filled grains per panicle, seed-setting rate and 1000-grain weight of the mutant were significantly affected. Genetic analysis showed that the mutation was controlled by a single recessive gene, tentatively named lbsl1(t), which was mapped to the short arm of chromosome 6. By developing simple sequence repeat (SSR) markers, the gene was finally delimited to an interval of 130 kb between markers RM586 and RM588. The lbsl1(t) gene is likely a novel rice spotted-leaf gene since no other similar genes have been identified near the chromosomal region. The genetic data and recombination populations provided will facilitate further fine-mapping and cloning of the gene.     

水稻脆秆突变体bc1-wu3的鉴定与基因克隆

【目的】茎秆机械强度与植株抗倒伏性直接相关。发掘脆秆突变体,克隆其相关基因有助于了解茎秆机械强度遗传机制,为抗倒伏育种提供理论依据。【方法】在经~(60)Co-γ诱变的粳稻品种武育粳3号后代群体中获得一个脆秆突变体,命名为bc1-wu3(brittle culm 1 from Wuyujing3),以突变体bc1-wu3为母本,Kasalath为父本构建相应的F_2分离群体,采用图位克隆的方法定位相应脆秆基因。【结果】与野生型相比,突变体的叶片、叶鞘、茎秆等组织在全生育期始终表现为脆性,易折断;穗长和粒长显著降低,粒宽显著增加。茎秆细胞细胞壁糖成分测定表明,细胞壁中纤维素含量极显著下降,而木糖、葡萄糖及阿拉伯糖含量则显著增加。茎秆切片观察发现突变体茎秆的厚壁细胞层数减少,厚壁细胞细胞壁极显著变薄。遗传分析表明,bc1-wu3脆性性状受1对隐性核基因控制。采用图位克隆的技术将该基因定位于第3染色体标记MK12与MK18之间,物理距离为57 kb,在此定位区段内包含1个已克隆的脆性基因BC1(LOC_Os03g30250)。测序结果表明,突变体bc1-wu3中BC1基因第2外显子内(CDS 659处)有1个碱基的替换(G-T),导致编码氨基酸由半胱氨酸变异为苯丙氨酸。实时荧光定量PCR结果表明,BC1基因在脆秆突变体bc1-wu3茎秆中的表达量显著降低。【结论】据此推断本研究中定位的脆秆基因bc1-wu3为BC1新等位基因。相关结果加深了对BC1基因功能的认识,有助于阐明水稻茎秆强度遗传机制。     

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。     

Dissection of QTLs for Hull Silicon Content on the Short Arm of Rice Chromosome 6

The QTL qHUS6 for hull silicon content in rice was previously located on the short arm of rice chromosome 6.By using an F2:3 population segregating in the RM587-RM19784 region harboring qHUS6 in an isogenic background,two QTLs for hull silicon content were detected,of which qHUS6-1 was located in the distal region and qHUS6-2 in the region proximal to the centromere.Three rice plants carrying small heterozygous segments in the target region were selected,of which two covered the qHUS6-1 region and the other...     

水稻黄绿叶突变体djyg的遗传分析与基因定位

在粳稻品种Dongjin大田种植过程中,发现一个黄绿叶自然突变体,命名为djyg。该突变体在苗期表现明显的黄绿叶表型,抽穗以后,叶色逐渐恢复正常。叶绿素含量测定结果表明,在苗期、分蘖盛期及抽穗期叶绿素b的含量分别下降53%、62%、36%。电镜结果表明,分蘖期突变体中基粒、类囊体垛堆凌乱、排列疏松,类囊体基质较为稀薄。qRT-PCR结果证实,PORA、Cab1R、PsbA的表达量在突变体中均较野生型明显下调。遗传分析结果表明,黄绿叶突变体djyg由一对隐性主效核基因控制,图位克隆确定该候选基因为编码叶绿素合成酶基因YGL1的一个新等位基因。该突变体未影响植株的主要农艺性状,可作为一个理想的表型标记应用于杂交稻育种工作中。     

水稻半矮杆突变体‘双辐矮糯’的农艺性状与遗传分析

倒伏是影响水稻（Oryza sativa L.）生产的重要因素,半矮杆是协调高产和抗倒性的理想资源。前期通过辐射诱变‘湘辐糯1号’获得了半矮杆突变体‘双辐矮糯’。本研究分析了‘双辐矮糯’的茎节配置特点、基本农艺性状及其遗传规律。结果表明：‘双辐矮糯’株高较‘湘辐糯1号’（133.13 cm）降低了24.88 cm,穗、第1节和第2节长度显著缩短,降幅分别达到18.10%、23.06%和19.13%,而第3、4、5节长度缩短程度不明显,表明其株高突变基因主要控制穗和高位茎节的伸长。‘双辐矮糯’的单株有效穗为10.4个,是‘湘辐糯1号’（5.2个）的2.04倍,其强分蘖能力确保了在每穗总粒数下降20.24%和千粒重下降7.51%情况下单株产量仍表现出25.26%的增长。品质分析发现,‘双辐矮糯’的粒长、粒宽、长宽比等外观品质指标及直链淀粉含量、胶稠度等食味品质指标与‘湘辐糯1号’无显著差异,说明‘双辐矮糯’株高的下降未对外观品质与食味品质产生不利影响。遗传分析发现,48对SSR标记在‘双辐矮糯’与‘湘辐糯1号’间扩增出了完全一致的基因型,二者遗传背景相似度达到了100.00%,证实了‘双辐矮糯’是源于‘湘辐糯1号’的真实变异。利用‘双辐矮糯’与‘湘辐糯1号’构建了2个正反交F₂群体,分析发现群体中半矮杆植株与野生型植株理论分离比符合1∶3,表明半矮杆性状受一个隐性细胞核基因控制。本研究明确了‘双辐矮糯’的株高表型和遗传特点,为深入挖掘其育种价值奠定基础。     

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。     

Tagging of Brown Planthopper Resistance Genes in F2s of IR50 × Ptb33 of Rice by Using Bulked Segregant Analysis

Brown planthopper(Nilaparvata lugens St?l) is one of the most damaging pests causing hopper burn in rice,and thereby reducing the productivity and also the quality of the product.The effective management strategy to control this pest is the identification and transfer of desirable genes to local rice cultivars.The most important approach for developing resistant cultivars is the identification of markers,which can help in marker-assisted selection of more durable resistant genotype.The susceptible parent IR50 and the resistant parent Ptb33,and their F2 populations were used in bulked segregant analysis for identification of resistant genes with random amplified polymorphic DNA marker(RAPD) primers.The primers OPC7 and OPAG14 showed both dominant and susceptible specific banding pattern so called co-dominant markers.Moreover,OPC7697 and OPAG14680 showed resistant specific bands and thus being in coupling phase,whereas OPC7846 and OPAG14650 showed susceptible specific genotypic bands in bulked segregant analysis.Therefore,the coupling phase markers,OPC7697 and OPAG14680,are considered to be more useful in marker-assisted selection of rice genotypes in crop improvement.     

水稻类树状突变体pla1-5的鉴定与基因克隆

在粳稻品种台北309经60 Co-γ辐射诱变的后代中发现了一份类树状突变体pla1-5,该突变体表现为植株矮化、叶片小且数量增多、分蘖数减少、高位分蘖、穗分化受阻等特征。遗传分析表明,该性状受一对隐性核基因控制。利用图位克隆技术将目的基因定位在第10染色体长臂上两个分子标记CHR1027和CHR1030之间,物理距离为58kb,并与SSR标记CHR1028和CHR1029共分离。根据水稻基因组序列的注释,该区域内存在5个完整的预测基因。测序分析表明pla1-5突变体中一个编码细胞色素P450CYP78A11的释义基因LOC_Os10g26340第1外显子内缺失了一个碱基T,导致移码突变和翻译提前终止。据此,初步推测细胞色素P450CYP78A11基因为PLA1-5的候选基因。     

水稻类病斑突变体lmm7的鉴定与基因定位

【目的】对水稻类病斑突变体的研究有助于解析其与植物生长和防御反应的关系。【方法】本研究在粳稻品系FI135胚培养过程中获得了1个类病斑突变体lmm7(lesion mimic mutant 7)。通过对其进行系统的表型鉴定、农艺性状考查、超微结构观察、生理学特性分析,阐明LMM7基因对植物生长的调控。通过病原菌抗性鉴定,明确lmm7对植物防御反应的影响。利用9311B与突变体lmm7杂交所得F₂群体对该突变体进行了遗传分析和基因精细定位。【结果】该突变体苗期表型正常,分蘖初期,植株基部叶片从叶尖开始不断出现褐色斑点,并向整株扩散,且斑点数目随植株生长不断增加。与野生型相比,突变体的株高、穗长、有效穗数、每穗粒数、结实率及剑叶长宽都显著降低,但籽粒性状和抽穗期没有显著性差异。遮光处理表明,突变体lmm7的表型受到光照诱导,抽穗期突变体lmm7叶肉细胞严重失绿,光合色素含量显著降低。组织化学分析表明,突变体病斑处的H₂O₂含量显著升高。透射电镜观察结果表明,突变体lmm7叶肉细胞的叶绿体数目减少,叶绿体类囊体片层结构严重受损,细胞器肿胀解体,并出现大量嗜锇小体,同时病斑内部和周围区域积累了大量的ROS。抗性鉴定结果显示突变体lmm7稻瘟病抗性水平显著高于野生型。遗传分析表明lmm7的突变表型受单个隐性基因控制。利用图位克隆的方法,目的基因被定位于水稻第7染色体短臂两InDel标记7B35和7B43之间,区间范围约260 kb。测序结果表明该区间内候选基因LOC_Os07g0203700第2891位碱基T发生了单碱基缺失,导致后续移码突变及翻译提前终止。【结论】lmm7与spl5互为等位基因,其突变抑制了植株的生长,同时增强了对稻瘟病的抗性。     

一种水稻微效QTL精细定位和克隆新途径

水稻重要农艺性状一般由少数主效QTL和大量微效QTL共同控制.水稻主效QTL克隆已取得显著进展,而微效QTL由于遗传作用弱,表型鉴定易受测量误差影响,克隆进展缓慢,但微效QTL在水稻重要农艺性状调控中的作用不容忽视.本文介绍了一种水稻微效QTL精细定位和克隆的新途径.该途径包含2个阶段:1)应用剩余杂合体构建近等基因系...     

水稻淡褐斑叶突变体lbsl1的遗传分析与基因定位

 通过EMS诱变籼稻品种IR64获得一个稳定遗传的淡褐色斑点叶突变体lbsl1（light brown spotted leaf 1）。在自然条件下,突变体播种后10~14 d,叶片上出现淡褐色斑点,随后逐渐扩散至全叶,第1叶至剑叶上均有淡褐色斑,为全生育期性状。斑点性状的表达对株高、生育期、结实率和千粒重等农艺性状具有显著的影响。遗传分析结果表明,该淡褐色斑点叶性状受一个隐性核基因控制。将突变体lbsl1与正常叶色水稻Morobereken杂交构建F2定位群体,利用SSR标记,最终将该淡褐叶基因lbsl1(t)定位在第6染色体短臂上一个约130 kb的区段上。定位的结果和发展的群体为该基因的进一步精细定位和克隆奠定了基础。     

引发提高水稻劣变种子发芽率的蛋白质组学分析

引发处理是目前简便有效的提升种子发芽率和整齐度的方法,但引发提高水稻劣变种子发芽能力的机理并不清楚。利用蛋白质组学方法,分析清水引发和藤茶提取物二氢杨梅素引发2种处理方法与对照的差异蛋白,初步揭示引发提升种子发芽能力的机制。分析双向电泳图谱差异获得2倍以上显著差异蛋白点79个,通过MALDI TOF/TOF MS质谱分析鉴定出74个蛋白,其中2种引发处理与未引发对照相比同步上调或下调的共有蛋白,即引发相关蛋白有57个。分析引发相关蛋白可知,绝大多数的逆境防御蛋白类、能量相关蛋白类以及蛋白质合成和目标类蛋白丰度在引发处理中显著增加,推测引发提高种子发芽率的原因与逆境记忆、能量代谢活动和蛋白质合成能力增强等密切相关。     

High-Yielding Performance of A New Rice Variety,Ir53650In Mildly Improved Acid Sulfate Soil Conditions

Abstract

A spectral and polarization image observation technique for detecting multiband polarimetric characteristics of reflected light from field-growing plants under daylight conditions was developed and the potential application of the method to in-situ assessments of wheat-leaf orientation at the heading stage was assessed. The developed digital imaging system corresponded to wavelength bands centered at 470, 550, and 647 nm, each with bandwidths of 10 nm. The instrument was fitted with a glass polarizer, which rotated from 0 to 360º in 15º steps, and polarized images of 1360 ×1024 pixels were captured at heading in wheat plots subjected to different fertilizer regimes at the jointing stage. Degree of polarization (DP) and mean brightness (MB) of the three bands were calculated from images of several pairs of top-dressed and non-top-dressed (non-dressed) plots, with a camera depression angle of 15–20º on two clear days. The relative azimuth angles between the view and insolation were approximately 135º (oblique front) and 180º (right in front), respectively. The mean DP for each plot area in the images varied between 0.3 and 1.4%. Although most of the top-dressed plots had significantly higher DPs than the non-dressed plots in the 550 nm band, few of the MB images in any band showed a clear difference between the top-dressed and non-dressed plots.