水稻分蘖的激素调控研究进展

水稻分蘖受外界环境和内源激素的影响,生长素、细胞分裂素和独角金内酯是目前已知影响水稻分蘖的3种主要植物激素。综述了这3种激素在调控水稻分蘖过程中的作用及相关的信号途径。     

水稻茉莉素受体基因OsCOI1a的克隆与功能研究

将拟南芥茉莉素受体基因COI1在水稻中的同源基因OsCOI1a构建到植物双元表达载体pMYC2,导入拟南芥茉莉素受体突变体coi1-1中,采用Western blot检测OsCOI1a基因的蛋白表达水平,通过观察突变体花粉的活性和果荚的育性,检测OsCOI1a对coi1-1突变体的互补情况,研究OsCOI1a基因的功能。结果表明,OsCOI1a基因能够在拟南芥coi1-1突变体中正确表达,并能部分互补雄性不育的表型,表明OsCOI1a基因在水稻茉莉素信号通路中可能起茉莉素受体作用。     

一份水稻寡分蘖近等基因系的构建及全基因组差异表达分析

利用水稻寡分蘖突变体G069与广亲和材料02428杂交和连续回交,建立了寡分蘖基因在02428基因组背景下的近等基因系02428 ft1.在分蘖始期对其分蘖节位的石蜡切片观察表明,寡分蘖基因抑制了水稻侧芽的发生。利用含有59 712个预测基因或已知基因的表达特征序列的基因芯片进行全基因组表达谱分析,结果表明共有136个cDNA克隆的表达在近等基因系02428 ft1中发生了变化,其中27个被激活,30个沉默,受到了寡分蘖基因的明显调控。功能分析表明, 这些受寡分蘖基因调控的基因功能分布较为广泛,包括转录因子、激酶、功能蛋白和调节蛋白等,为水稻分蘖调控机制的研究提供了相关信息。     

水稻ABC1基因家族的鉴定及在非生物胁迫下的表达分析

ABC1(Activity of bc1 complex)家族属于蛋白质激酶家族,其成员普遍存在于原核和真核生物中.已有研究表明,几个植物ABC1基因参与非生物胁迫应答.为了解ABC1基因在水稻中的结构和功能,采用生物信息学方法分别在水稻和拟南芥上鉴定出15个和17个ABC1基因,并进行了系统发育和表达分析.结果表明,该家族在单、双子叶植物分离之前就已经发生了分化,其基本特征已经形成;单、双子叶植物分离之后,该家族在水稻和拟南芥中均以物种特异的方式进行了扩增.内含子/外显子结构分析显示多数直系同源基因之间外显子大小接近,而内含子差别较大,水稻含有更多大的内含子;内含子获得是近期伴随水稻ABC1家族进化的重要事件.多序列比对显示,ABC1结构域具有1个保守的氨基酸片段和4个保守的氨基酸残基.在线亚细胞定位预测9个水稻ABC1蛋白定位在叶绿体上.实时定量RT-PCR分析表明,水稻ABC1基因主要在叶片中表达,并且受多种非生物胁迫因素包括H2O2、脱落酸、低温、干旱、黑暗和高盐的调控.说明水稻ABC1家族不仅在逆境胁迫应答中发挥重要作用,可能还与水稻特定的生理过程有关.     

大豆GmWRI1a基因启动子克隆及其功能分析

为了探究大豆种子油脂合成与储存基因GmWRI1a的调节,分析其启动子功能,从大豆品种东农50基因组中扩增获得GmWRI1a基因转录起始位点上游1669bp的启动子序列,转化拟南芥。GUS染色确定了启动子的有效性,继而根据顺式作用元件的分布,分别扩增得到4个启动子5'端缺失片段1138bp,1087bp,690bp和437bp。4个启动子缺失片段分别转化拟南芥后,通过GUS组织染色结果发现,这些启动子片段能够驱动下游GUS基因表达。GUS酶活表明,启动子存在乙烯、茉莉酸、赤霉素3种激素响应元件,其中乙烯、茉莉酸和赤霉素的响应元件分别分布在-1138^-1087bp、-1087^-690bp和-690^-437bp。     

水稻ABC1基因家族的鉴定及在非生物胁迫下的表达分析

 ABC1（Activity of bc1 complex）家族属于蛋白质激酶家族,其成员普遍存在于原核和真核生物中。已有研究表明,几个植物ABC1基因参与非生物胁迫应答。为了解ABC1基因在水稻中的结构和功能,采用生物信息学方法分别在水稻和拟南芥上鉴定出15个和17个ABC1基因,并进行了系统发育和表达分析。结果表明,该家族在单、双子叶植物分离之前就已经发生了分化,其基本特征已经形成;单、双子叶植物分离之后,该家族在水稻和拟南芥中均以物种特异的方式进行了扩增。内含子/外显子结构分析显示多数直系同源基因之间外显子大小接近,而内含子差别较大,水稻含有更多大的内含子;内含子获得是近期伴随水稻ABC1家族进化的重要事件。多序列比对显示,ABC1结构域具有1个保守的氨基酸片段和4个保守的氨基酸残基。在线亚细胞定位预测9个水稻ABC1蛋白定位在叶绿体上。实时定量RT-PCR分析表明,水稻ABC1基因主要在叶片中表达,并且受多种非生物胁迫因素包括H₂O₂、脱落酸、低温、干旱、黑暗和高盐的调控。说明水稻ABC1家族不仅在逆境胁迫应答中发挥重要作用,可能还与水稻特定的生理过程有关。
     

拟南芥AtLURP1基因启动子在转基因烟草和水稻中的功能分析

克隆拟南芥LURP1基因上游1515 bp的启动子调控序列并命名为AtLURP1p,将其与GUS报告基因融合,构建成植物表达载体,分别遗传转化烟草和水稻,获得LURP1p::GUS的烟草和水稻转基因植株及其相应的T2代株系,分别研究LURP1p对水稻稻瘟病、辣椒青枯病侵染及SA、MeJA、ABA等几种重要的植物激素信号分子处理的应答.结果表明:(1)转基因烟草和水稻在几种激素,包括SA、MeJA、ABA的诱导处理下,GUS基因均可以被诱导表达;(2)转基因烟草在细菌性病菌青枯病的侵染下,GUS可被诱导表达并表现出持续表达的趋势,转基因水稻在稻瘟病的侵染下,其GUS基因也被诱导表达,并表现出后期持续上调的趋势.这些研究结果表明,拟南芥LURP1的应答逆境信号通路也存在于烟草和水稻等植物,该启动子可用作诱导型启动子广泛地应用于不同植物抗病基因工程.     

大豆E3泛素连接酶基因GmHOS1a和GmHOS1b生物信息学分析和敲除载体构建

HIGH EXPRESSION OF OSMOTICALLLY RESPONSIVE GENES 1(HOS1)是植物中多种信号途径的整合因子,在植物发育、胁迫反应、光形态建成和开花调控中发挥至关重要的作用,是很有潜力的作物工程育种目标基因,但其在大豆中的功能尚未被揭示。为了揭示其在大豆中的功能,本研究利用反向遗传学,成功克隆了两个大豆HOS1基因,GmHOS1a和GmHOS1b,并通过生物信息学技术,对其蛋白结构、表达模式和功能进行了分析。进化树和结构域分析表明,GmHOS1a和GmHOS1b具有N端的环指结构域和与ELYS高度相似的保守基序,与拟南芥和水稻HOS1蛋白高度同源。亚细胞定位结果显示,GmHOS1a和GmHOS1b定位在细胞核中。Quantitative Real-time PCR(qRT-PCR)结果表明,GmHOS1a和GmHOS1b的基因表达模式相似,均在三出复叶中高度表达。48 h光周期RNA-sequencing(RNA-seq)和qRT-PCR分析表明,GmHOS1a和GmHOS1b基因的表达具有生物钟昼夜节律性,其中GmHOS1a的表达量显著高于GmHOS...     

甘蔗ScHTD2基因的克隆及生物信息学分析

D14是一类能够有效抑制水稻分蘖的水解酯酶,可能是独脚金内酯信号传导途径中的重要组分。为研究该基因在甘蔗分蘖性状中的功能,利用RT-PCR和RACE技术从甘蔗主栽品种ROC22中克隆出D14同源基因的c DNA全长,命名为Sc HTD2,并对其进行生物信息学分析。结果表明:Sc HTD2基因的c DNA全长为1 302 bp(KP137674.1),具有927 bp的开放阅读框,编码308个氨基酸;蛋白质分析表明,Sc HTD2为不稳定的水溶性非分泌蛋白,主要作用于氨基酸的生物合成或者作为生长因子调控生物生长发育。亚细胞定位于叶绿体,存在14个磷酸化位点,不存在乙酰化位点和信号肽。二级结构以α螺旋和无规则卷曲为主,还有一些残基形成β-折叠和链延伸。保守结构域和三级结构预测均表明该蛋白包含一个α保守水解酶家族,属"D14-Like"或者"DAD2-Like"蛋白家族。推测Sc HTD2可能作为独脚金内酯调控甘蔗分蘖信号转导中一个具有催化特性的水解脂酶而起作用。进化树分析表明该蛋白与高粱、玉米等单子叶植物进化关系较近。     

籼稻矮化多分蘖突变体gsor23的基因克隆与表达分析

 矮化多分蘖突变体gsor23是籼稻品种Indica9经γ射线辐射后获得的。遗传分析表明其矮化多分蘖表型受一个隐性基因控制,并将突变基因定位在第1染色体长臂上InDel标记C1 WT2与C1 WT4之间。这两个标记之间物理距离为386 kb,其间包含一个抑制植物分枝的基因D10。对gsor23中的D10基因测序发现编码区第404位的碱基C缺失,导致从第135位氨基酸开始移码突变。gsor23的突变位点与已报道的粳稻背景的d10 1和d10 2突变体不一样,是D10基因一个新的等位突变体。D10编码的类胡萝卜素裂解双加氧酶8 (carotenoid cleaving dioxygenase, CCD8),是抑制分枝的新型植物激素独脚金内酯(strigolactones, SLs)合成途径中的关键酶之一。用SLs人工合成类似物GR24处理gsor23,其多分蘖表型受到抑制。实时 RT PCR结果显示D10在水稻根部表达量较高,叶片较低。在突变体gsor23中,参与 SLs合成的基因D10上调表达,而可能参与SLs信号转导的基因D3和D14表达下调。     

Identification and Cloning of Tillering-Related Genes OsMAX1 in Rice

Tillering is an important agronomic trait which has a direct impact on plant type and grain yield. Strigolactones are a class of important phytohormones regulating rice tillering. ATMAX1 is an important gene involved in strigolactone biosynthesis through encoding the protein P450 in Arabidopsis. Based on sequence BLASTp, we identified five homologous genes of ATMAX1 in rice, i.e., OsMAX1a, OsMAX1b, OsMAX1c, OsMAX1d and OsMAX1e. Among them, OsMAX1a and OsMAX1e showed stable and high expression in rice tissues. In addition, we observed that Os MAX1a and OsMAX1e can rescue the branched phenotype and the influences caused by MAX1 mutation in Arabidopsis. Moreover, the expression of OsMAX1a and OsMAX1e can respond to phosphate deficiency and different phytohormones, especially GR24, a strigolactone analogue. Therefore, it is concluded that Os MAX1a and OsMAX1e are involved in the biosynthesis of strigolactones and regulated rice tillering.     

Mixing properties and dough functionality of transgenic lines of a commercial wheat cultivar expressing the 1Ax1, 1Dx5 and 1Dy10 HMW glutenin subunit genes

In this work we report the effects of the HMW-GS 1Ax1, 1Dx5 and 1Dy10 on the breadmaking quality of the bread wheat cultivar Anza that contains the HMW-GS pairs 1Dx2 + 1Dy12 and 1Bx7* + 1By8, and is null for the Glu-A1 locus. This allows the characterization of individual subunits 1Dx5 and 1Dy10 in the absence of subunit 1Dx5, and the interactions between these subunits and subunits 1Dx2 and 1Dy12 to be determined. Three transgenic lines termed T580, T581 and T590, containing, respectively, the HMW-GS 1Ax1, 1Dx5 and 1Dy10 were characterized over 3 years using a range of widely-used grain and dough testing methods. The transgenic subunits 1Ax1, 1Dx5 and 1Dy10 accounted for 25.2%, 20.3% and 17.9%, respectively, of the total HMW-GS in the three transgenic lines. Although lines T581 and T590 expressed similar levels of subunits 1Dx5 and 1Dy10 they had different effects on other aspects of protein composition, including changes in the ratios of glutenin/gliadin, of HMW/LMW-GS, the 1Dx2/1Dy12, the x-type/y-type HMW-GS and the proportions of high molecular mass glutenin polymers. In contrast, lines transformed to express subunits 1Ax1 and 1Dx5 showed similar changes in protein composition, with higher protein contents and decreased ratios of glutenin/gliadin and 1Dx2/1Dy12. In addition, both transgenic lines showed similar increases in the ratio of x-type/y-type subunits compared to the control line. The transgenic lines were analysed using Farinograph, Mixograph and Alveograph. This confirmed that the expression of all three subunits resulted in increased dough strength (and hence breadmaking quality) of the cultivar Anza. A beneficial effect of subunit 1Dx5 has not been reported previously, transgenic wheat lines expressing this subunit giving overstrong dough unsuitable for breadmaking. However, the expression of subunit 1Dy10 had a greater effect on breadmaking quality than subunits 1Ax1 and 1Dx5. The Farinograph parameters such as dough stability and peak time were increased by 9.2-fold and 2.4-fold, respectively, in line T590 (expressing 1Dy10) with respect to the control line. Similarly, the Mixograph mixing time was increased by four-fold and the resistance breakdown decreased by two-fold in line T590 compared with the control line. The Alveograph W value was also increased by 2.7-fold in line T590 compared to the control line. These transgenic lines are of value for studying the contribution of specific HMW-GS to wheat flour functional properties.     

Effects of genes increasing the number of spikelets per panicle,TAW1 and APO1, on yield and yield-related traits in rice

The genes TAWAWA1 (TAW1) and ABERRANT PANICLE ORGANIZATION1 (APO1) increase the number of spikelets per panicle (SN). In the present study, we examined the effects of these genes on morphological traits, yield, and yield-related traits including yield components using the near-isogenic lines (NILs) in the genetic background of a japonica rice variety, Koshihikari – NIL-taw1, NIL-apo1-D3, and NIL-apo1-D4 – in a field experiment. The SN and total number of spikelets per area of the three NILs were larger than those of Koshihikari. However, the yield of the three NILs did not exceed that of Koshihikari due to their low filling ability. Interestingly, our field experiments indicated that TAW1 did not affect the diameter of internodes and the PN, whereas APO1 decreased the PN and increased the diameter of internodes. These results suggest that TAW1 and APO1 differently affect yield-related traits.     

Omega-3 Fatty Acids Upregulate SIRT1/3, Activate PGC-1α via Deacetylation,and Induce Nrf1 Production in 5/6 Nephrectomy Rat Model

Mitochondrial dysfunction contributes to the pathogenesis of kidney injury related with cardiovascular disease. Peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) protects renal tubular cells by upregulating nuclear factor erythroid 2-related factor 2 (Nrf2). AMP-activated protein kinase (pAMPK)-mediated phosphorylation and sirtuin 1/3 (SIRT1/3)-mediated deacetylation are required for PGC-1α activation. In the present study, we aimed to investigate whether omega-3 fatty acids (FAs) regulate the expression of mediators of mitochondrial biogenesis in 5/6 nephrectomy (Nx) rats. Male Sprague-Dawley rats were assigned to the following groups: sham control, Nx, and Nx treated with omega-3 FA. The expression of PGC-1α, phosphorylated PGC-1α (pPGC-1α), acetylated PGC-1α, and factors related to mitochondrial biogenesis was examined through Western blot analysis. Compared to the control group, the expression of PGC-1α, pAMPK, SIRT1/3, Nrf1, mTOR, and Nrf2 was significantly downregulated, and that of Keap 1, acetylated PGC-1α, and FoxO1/3, was significantly upregulated in the Nx group. These changes in protein expression were rescued in the omega-3 FA group. However, the expression of pPGC-1α was similar among the three groups. Omega-3 FAs may involve mitochondrial biogenesis by upregulating Nrf1 and Nrf2. This protective mechanism might be attributed to the increased expression and deacetylation of PGC-1α, which was triggered by SIRT1/3.     

甲基异柳磷防治花生根结线虫病的研究

１９８３—１９９１年用杀虫剂甲基异柳磷对花生根结线虫病的防效进行了研究，结果表明，于花生播种时，按每公顷用４０％甲基异柳磷乳油有效成分４．５ｋｇ，兑土或水稀释后，施于播种沟内，防效为４６．１％，增产７５％，并有兼治种苗期蚜虫等多种害虫作用。适合于花生根结线虫病发生较轻而虫害发生严重的地区或田块施用。     

宁夏引黄灌区冬小麦谷蛋白亚基组成和1BL/1RS易位分析及品质性状研究

为全面了解宁夏引黄灌区冬小麦品质概况,为冬小麦品质改良和粮食生产提供理论依据。选用冬小麦主栽品种、亲本材料和高代品系30份。用SDS-PAGE分析了其中18份材料的HMW—GS、LMW—GS组成和1BL／1RS易位系分布状况。并对23个冬小麦品种的营养与加工品质进行了研究。结果表明,2#、7＋9、2＋12、Glu-A3a、Glu-A3c、Glu-B3h和Glu—B3j在宁夏引黄灌区冬小麦中分布较广,1BL／1RS易位系分布相当普遍。分布频率为27．8％。参试品种的籽粒硬度、面粉PPO活性、SDS沉淀值、形成时间、稳定时间和评价值的变异范围较大,变异系数分别为25．37％、33．68％、21．42％、26．58％、43．95％和29．31％。多数冬小麦品种（系）的千粒重、出粉率和湿面筋含量低于对照宁春4号。而蛋白质含量、SDS沉淀值、吸水率和稳定时间优于宁春4号。供试品种（系）中。HMW-GS品质评分、籽粒硬度、蛋白质含量、SDS沉淀值均较高,并且不含1BL／1RS易位系的有烟优361、济麦20、鲁875067和923—9等,可用于宁夏引黄灌区冬麦品质改良。     

非1B/1R和1B/1R 类型小麦K型雄性不育系穗长和小穗数性状的遗传分析

为明确非1B/1R和1B/1R 类型小麦K 型雄性不育系穗长和小穗数的遗传规律,利用非1B/1R类型KTSP732A和1B/1R类型K3314A 两种K型不育系材料,采用主基因+多基因混合遗传模型分析法,对穗长和小穗数两类性状进行单世代和多世代混合分离分析.结果显示,非1B/1R类型KTSP732A和1B/1R类型K3314A穗长性状均符合加性-显性-上位性多基因模型(C-0),无主基因存在;非1B/1R类型KTSP732A的小穗数性状符合两对等显性主基因+加性-显性多基因混合模型(E-6),主基因遗传率为40.7%;而1B/1R类型K3314A的控制小穗数性状基因类型与非1B/1R不同,该类型无主基因存在,也符合加性-显性-上位性多基因模型(C-0).     

黑粒小麦漯珍1号的C-分带及GISH和PAGE分析

为了解黑粒小麦漯珍1号的染色体构成、蛋白组分以及抗病性,利用染色体C-分带、基因组原位杂交(GISH)和PAGE分析对漯珍1号进行了系统鉴定。结果表明,漯珍1号含1对1BL/1RS易位染色体,另外,其7B染色体的C-分带带型也表现出与中国春有较大差异,而其余染色体则未见明显区别。其PMC M染色体平均构型为1.97(棒状二价体) 19.03(环状二价体),说明具有较好的细胞学稳定性。SDS-PAGE分析表明,其高分子量谷蛋白亚基(HMW-GS)组成为1、7 8和5 10;种子醇溶蛋白A-PAGE鉴定显示,除含有黑麦1RS GldB3编码的醇溶蛋白外,还有许多迁移率明显不同于中国春的多态性位点。抗病性初步鉴定结果表明,漯珍1号高感赤霉病和白粉病。     

GmHs1~(pro-1)多样性分析及蛋白互作预测

GmHs1~(pro-1)是Hs1~(pro-1)同源的抗线虫候选基因,通过使用不同的大豆品种克隆GmHs1~(pro-1)CDS区研究了其多样性、建立了进化树,并利用在线数据库分析预测了与其互作的蛋白。结果表明:在24种大豆品种中均克隆到长度约1 400bp的GmHs1~(pro-1)CDS区,同已有数据合并后该区段共有20个突变点,5个新突变点中的3个引起了编码氨基酸的变化。GmHs1~(pro-1)可能主要同glyma18g04770编码的蛋白互作,该蛋白受syringolide诱导。GmHs1~(pro-1)定位在胞浆内,具有1个吲哚化合物双加氧酶类似结构域,可能参与吲哚相关通路影响线虫发育。     

花生品种莆花1号丰产性、稳定性及适应性分析

以2006~2007年度福建省春花生区域试验结果为材料,通过产量、变异系数和回归系数分别对花生品种莆花1号和泉花10号的丰产性、稳定性和适应性进行分析与比较,以促进该品种在生产上的推广应用。结果表明,莆花1号的丰产性、稳定性和适应性优于对照品种,是一个比较理想的高产花生新品种。