不同类型水稻不育系药隔发育过程中Ca2+的分布变化

采用焦锑酸钾沉淀法研究了不同类型水稻不育系与其相应保持系及其可育花药药隔发育过程中Ca^2＋的分布变化。结果显示在正常花药的发育过程中,药隔中的Ca^2＋呈以下变化规律：（1）花药发育早期药隔中的Ca^2＋沉淀很少;（2）随着花药的发育,药隔中的Ca^2＋沉淀增加,木质部细胞的次生加厚壁上有较多的Ca^2＋沉淀,连接组织中的Ca^2＋沉淀也大大增加;（3）到了花粉内容物充实后期及成熟花粉时期,药隔中的Ca^2＋又略有减少。而不育花药药隔中的Ca^2＋在花药发育的各时期均比可育花药要多。败育类型不同,药隔中出现大量Ca^2＋沉淀的时期也不同,无花粉型不育系在花粉母细胞时期药隔中就有大量Ca^2＋沉淀,而农垦58S从单核花粉时期约隔中才出现大量Ca^2＋沉淀。实验结果表明：Ca^2＋在不同类型的不育水稻中呈现出一定的规律性变化,药隔中Ca^2＋的异常分布可能与水稻花粉的败育有关。     

热胁迫对辣椒花药发育过程中Ca2+分布的影响

运用焦锑酸钾沉淀法,研究了不同热胁迫时间对辣椒（Capsicum annuum L.)小孢子发生和花粉发育过程中Ca^2 分布的影响。在小孢子母细胞中,细胞表面有少量Ca^2 分布,细胞核中基本上观察不到Ca^2 ,热胁迫12h后,细胞质和细胞核中Ca^2 明显增多,液泡膜内侧也有许多Ca^2 分布,热胁迫24h后,大量的Ca^2 分布在细胞质中,液泡膜上和液泡内;在四分体时期,与小孢子母细胞相比,四分小孢子表面和细胞质中Ca^2 数量明显增加,热胁迫24h后,细胞质和细胞核中Ca^2 更多;在小孢子中,大量Ca^2 分布在壁上,质膜内侧,液泡膜上,少量分布在细胞质和细胞核中,热胁迫12h后,质膜上Ca^2 增多,24h后,细胞质,细胞核中,质膜内侧的Ca^2 继续增多,热胁迫对成熟花粉中Ca^2 的分布无明显影响。     

水稻胞质雄性不育系珍汕97A及其保持系珍汕97B绒毡层发育过程中的Ca2 +分布变化

利用焦锑酸钾沉淀法研究了野败不育系珍汕97A及其保持系珍汕97B绒毡层细胞的发育过程及其细胞中Ca2 +的分布变化。研究发现保持系绒毡层细胞在单核花粉晚期才开始迅速解体,而不育系绒毡层细胞在花粉母细胞时期就开始出现核膜、细胞膜解体,此过程持续到二核花粉时期。珍汕97A绒毡层细胞从花粉母细胞时期开始,细胞质内有少量颗粒状的Ca2 +沉淀;减数分裂时期,绒毡层细胞的内切向壁表面有大量大颗粒的Ca2 +沉淀;单核花粉时期绒毡层细胞周围集聚一层Ca2 +沉淀。而保持系绒毡层细胞遮花粉母细胞时期和减数分裂时期细胞内没有Ca2 +沉淀;单核花粉时期绒毡层细胞内的Ca2 +沉淀主要分布在解体的细胞质内。推测绒毡层细胞结构发育的异常和Ca2 +的异常分布可能与花粉的败育有关。     

水稻胞质雄性不育系珍汕97A及其保持系珍汕97B绒毡层发育过程中的Ca2+分布变化

利用焦锑酸钾沉淀法研究了野败不育系珍汕97A及其保持系珍汕97B绒毡层细胞的发育过程及其细胞中Ca2 的分布变化。研究发现保持系绒毡层细胞在单核花粉晚期才开始迅速解体,而不育系绒毡层细胞在花粉母细胞时期就开始出现核膜、细胞膜解体,此过程持续到二核花粉时期。珍汕97A绒毡层细胞从花粉母细胞时期开始,细胞质内有少量颗粒状的Ca2 沉淀;减数分裂时期,绒毡层细胞的内切向壁表面有大量大颗粒的Ca2 沉淀;单核花粉时期绒毡层细胞周围集聚一层Ca2 沉淀。而保持系绒毡层细胞遮花粉母细胞时期和减数分裂时期细胞内没有Ca2 沉淀;单核花粉时期绒毡层细胞内的Ca2 沉淀主要分布在解体的细胞质内。推测绒毡层细胞结构发育的异常和Ca2 的异常分布可能与花粉的败育有关。     

水稻细胞质雄性不育系及其保持系幼穗发育过程中内源激素的变化

利用酶联免疫测定技术研究了水稻细胞质雄性不育系珍汕97A及其保持系珍汕97B在幼穗发育过程中叶片、幼穗和花药中内源IAA、GA1＋4、ABA和iPAs含量的动态变化。结果表明,1）从幼穗发育的雌雄蕊形成期到三核花粉期,保持系叶片中IAA水平高于不育系,并在二核花粉期最高;在幼穗和花药中也以保持系为高。2）保持系与不育系叶片中GA1＋4含量变化趋势相似,均为先升后降,但从单核到三核花粉期以保持系为高;在幼穗和花药中也都以保持系为高。3）不育系叶片中ABA水平在幼穗发育早期明显高于保持系,中期与保持系相近,后期又高于保持系;在幼穗和花药中也都以不育系为高。4）保持系叶片、幼穗和花药中iPAs含量始终显著高于不育系。5）保持系叶片中IAA＋GA1＋4＋iPAs与ABA之比值也始终高于不育系。提示不育系叶片、幼穗和花药中IAA、GA1＋4、iPAs和ABA含量出现了异常,且IAA、GA1＋4和iPAs亏缺以及ABA盈积可能与水稻细胞质雄性不育发生有关。     

水稻细胞质雄性不育系及其保持系幼穗发育过程中内源激素？…

利用酶联免疫测定技术研究了水稻细胞质生不育系珍汕９７Ａ及其保持系珍汕９７Ｂ在幼穗发育过程中叶片,幼穗和花药中内源ＩＡＡ,ＧＡ１＋４,ＡＢＡ和ｉＰＡｓ含量的动态变化,结果表明：１）从穗发育的雌雄蕊形成期到三核花粉期,保持系叶片中ＩＡＡ水平高于不育系,并在二核花粉最高;在幼穗和花药中也可保持系为高。２）保持系与不育系叶片中ＧＡ１＋４含量变化趋势相似,均为先升后降,但从单核到三核花粉期以保持系为高;在粉     

水稻细胞质雄性不育系及其保持系幼穗发育过程中的多胺代谢

在幼穗发育过程中,不育系和保持系统穗多胺含量先剧降后稳定或略回升,精氨酸脱羧酶活性快速下降,而二胺和多胺氧化酶活性缓慢下降,从雌雄蕊形成期到花粉母细胞形成期,不育系的多胺含量和精氨酸脱羧酶活性明显低于保持系;不过,两系二胺氧化酶和多胺氧化酶活性却判别不大。     

云南紫稻细胞质雄性不育系花药发育过程中Ca2+的分布特点

运用焦锑酸钾沉淀法研究了云南紫稻细胞质雄性不育系和保持系花药在发育过程中Ca^2 的分布特点。结果表明,保持系的花粉母细胞和小孢子的胞质内部基本无Ca^2 的沉淀,后期花粉外壁出现Ca^2 的沉淀;保持系早期的绒毡层细胞形态正常,胞内有少量Ca^2 沉淀,后期绒毡层细胞开始凋亡,胞质凝集,胞内出现大量Ca^2 的颗粒。不育系花粉母细胞在减数分裂时期败育,胞质液泡化,内部出现大量Ca^2 的沉淀;不育系绒毡层细胞形态正常,胞内无Ca^2 的沉淀。绒毡层与花粉母细胞、小孢子之间出现大量Ca^2 颗粒。探讨了不育系花药花粉母细胞中以及与绒毡层细胞之间Ca^2 的异常积累与雄性不育的关系。     

红莲型水稻细胞质雄性不育花粉总蛋白质初步比较分析

采用固相pH梯度/SDS-PAGE双向电泳对红莲型细胞质雄性不育水稻的不育系(YTA)和保持系(YTB)二核期花粉总蛋白质进行了分离,通过银染显色,获得了分辨率和重复性较好的双向电泳图谱。用PDQuest2DE软件可识别约1500个蛋白质点,其中差异表达的蛋白质点数为120。将其中15个差异点采用基质辅助激光解析电离飞行时间质谱(matrix assisted laser desorption/ionizaton time of flight mass spectrometry,MALDI-TOF-MS)进行了肽质指纹图分析,通过采用Mascot软件对MSDB数据库查询,其中7个蛋白质点得到了鉴定。YTA相对于YTB有部分参与物质和能量代谢的蛋白质缺失或表达量降低,这些蛋白质分别是水稻线粒体H -转运ATPase(H -ATPase)α链、盐诱导型膜联蛋白、线粒体NAD -依赖型苹果酶和磷酸核糖焦磷酸合成酶等。这些蛋白质的表达下调或缺失可能与线粒体提供能量不足而导致的花粉不能正常发育有关。线粒体电压依赖性阴离子通道(VDAC)这一重要蛋白质在YTA中的上调表达有可能与花粉败育过程中细胞的程序性死亡相关。     

水稻细胞质雄性不育系与保持系的呼吸途径比较

比较水稻细胞质雄性不育系珍汕９７Ａ及其保持系珍汕９７Ｂ之幼德、叶片和花药的呼吸代谢。结果表明：不育系花药的总呼吸速率、抗氰呼吸所占总呼吸比较和细胞色素氧化酶（ＣＯＤ）、苹果酸脱氢酶（ＭＤＨ）和６－磷酸葡萄糖脱氢酶（６－ＰＧＤＨ）活性低于保持系;幼穗中则仅６－ＰＧＤＨ低于保持系;叶片间各指标均无差异。Ｎａ３ＰＯ４可抑制保持系的叶片和幼穗及不育系叶片总呼吸的３０％,而仅能抑制不育系幼穗总呼吸的２４％;     

水稻三系不育系育性与温度关系研究

通过自然条件下分期播种实验,对生产上应用的12个三系不育系花粉育性与温度的关系进行比较分析。结果表明:自然高温条件下不育系金23A、T98A、丰源A及冈46A花粉不育性稳定;隆398A、隆香634A、4302A、优IA及D62A出现少量可染花粉;高温诱导天丰A、五丰A和粤泰A花粉可育作用明显。此外,对可染花粉率与水稻不同发育时期的日均温相关性的分析表明,五丰A、天丰A和粤泰A可染花粉率均与抽穗前13~18 d的日均温显著相关(P0.05),高温诱导五丰A、天丰A和粤泰A育性转换敏感期为雌雄蕊原基形成末期至减数分裂期,育性转换临界日均温度分别为27.11℃、26.48℃和27.30℃。     

Mechanism of Anther Dehiscence in Rice (Oryza sativa L.)

This paper presents a new explanation of the mechanism of antherdehiscence in rice during the period from floret opening topollen dispersal. The theca dehisced on the stomium in the apicalpart and the anther wall in the basal part of the large locule.Comparison of the anther dehiscence process under various airhumidity conditions showed that the process, until the splittingat the apical and basal parts, was a moisture-requiring processwhereas the widening of the splits in both parts was a desiccatoryprocess. Observation of the anther transverse section, revealedthe marked development of the U-shaped thick cell wall in theendothecium adjacent to these two splits. From these observations,the anther dehiscence mechanism may be explained as follows.At the time of anthesis, pollen grains swell rapidly in responseto the floret opening and cause the theca to bulge, rupturingthe septum. The pollen pressure combined with the inward bendingof the locule walls adjacent to the stomium causes splittingof the stomium in the apical part of the theca. At the sametime, the septum rupture extends to the bottom of the largelocule supported by the pollen pressure. After these processes,the locule walls adjacent to both splits straighten probablydue to their water loss. This straightening widens the splitsand the swollen pollen grains overflow from the widened splits.Copyright1999 Annals of Botany Company Anther dehiscence, Oryza sativa L., pollen grain swelling, rice, septum, stomium, theca.     

Selection of Primary Trisomics from Anther Culture of Autotetraploid Rice (Oryza sativa L.)

The hybrids from different generations of autotetraploid rice (Oryza sativa L. ) and the original autotetraploid rice (indica and japonica) were used for anther culture, and the pollen-plantlets from them were induced. Due to the significant difference on phenotype among the trisomics and between trisomics and diploid, 15 lines of 4390 H1 induced plants were selected for chromosome study. Their PMC meiosis were observed. The results showed that the chromosomes from these plants consisted of 2n, 4n and aneuploids, and their ratios were 88.00%, 5.53% and 6.47% respectively. 272 trisomics from 284 aneuploids were identified, which acounted for 6.20% of all the pollen-plants. According to the special characters from the whole set of trisomics, they were classified as 9 types. The 9124- 7 trisomics were designated as triplo-8 by the pachytene analysis. Sowing the seeds of triplo8, the transmission rate of extra chromosome was calculated at the seedling stage of H2. The rate of trisomic was 34.11% of all plants, the agronomic characters were similar to the H1 parent plants.     

水稻花药培养力的遗传分析及基因定位

在栽培稻的籼粳亚种间,花药培养力存在显著差异,这一差异主要是由遗传因素引起的。以适合籼粳稻杂种花药培养的SK_3培养基,经花药培养获得了一个籼粳交F_1代的加倍单倍体(DH)群体,对该群体的110个株系用同一种培养基进行花药培养,利用该群体构建的分子图谱进行有关水稻花药培养力的数量性状基因座位(QTLs)的分析。结果表明,与水稻花药培养力有关的4个性状在DH群体中均表现为连续分布,愈伤组织诱导率与绿苗分化率之间不存在相关性,而绿苗产率与愈伤组织诱导率和绿苗分化率均显著相关。在第6、7、8、10和12 5条染色体上分别检测到与愈伤组织诱导率有关的5个QTLs,其加性效应均为正。在第1和第9染色体上检测到与绿苗分化率有关的2个QTLs,这两个性状间的QTs不存在连锁。在第9染色体上有一个主效基因与白苗分化率有关,对绿苗产率则没有检测到特有的QTL。     

水稻体细胞无性系变异

水稻体细胞无性系变异研究取得了很大进展 ,获得了大量抗病、抗逆、优质、矮杆等突变体。对这些突变体遗传分析表明 ,大多数突变性状由 1对或 2对基因控制。水稻体细胞无性系变异的发生与基因型、性状、继代时间、培养方式等有关 ,并具有内在的机制 ,点突变和反转录转座子插入可能是引起水稻无性系变异的两个重要原因。     

Coleoptile Senescence in Rice (Oryza sativa L.)

We investigated the cellular events associated with cell deathin the coleoptile of rice plants (Oryza sativa L.). Seeds germinatedunder submergence produced coleoptiles that were more elongatedthan those grown under aerobic conditions. Transfer of seedlingsto aerobic conditions was associated with coleoptile opening(i.e. splitting) due to death of specific cells in the sideof the organ. Another type of cell death occurred in the formationof lysigenous aerenchyma. Senescence of the coleoptile was alsonoted, during which discolouration of the chlorophyll and tissuebrowning were apparent. DNA fragmentation was observed by deoxynucleotidyltransferase-mediateddUTP nick end labelling (TUNEL) assay, and further confirmedby the appearance of oligonucleosomal DNA ladders in senescentcoleoptile cells. Two nucleases (Nuc-a and Nuc-b) were detectedby in-gel-assay from proteins isolated from coleoptiles. Nuc-a,commonly observed in three cell death phases required eitherCa²⁺or Mg²⁺, whereas Nuc-b which appeared during senescencerequired both Ca²⁺and Mg²⁺. Both nucleases were strongly inhibitedby Zn²⁺. Copyright 2000 Annals of Botany Company Aerenchyma, rice, cell death, coleoptile, fragmentation, nuclease, Oryza sativa, senescence, split, submergence, TUNEL     

Cold stress-induced calcium-dependent protein kinase(s) in rice (Oryza sativa L.) seedling stem tissues

Ca²⁺-dependent protein kinases (CDPKs) play an important role in plant signal transduction. Protein kinase(s) activities induced by 5°C cold stress in rice (Oryza sativa L.) seedlings were investigated in both leaf and stem tissues in an early (up to 45 min) and late (up to 12 h) response study. The leaf had 37-, 47- and 55-kDa protein kinase activities, and the stem had 37-, 47- and 55-kDa protein kinase activities. A 16-kDa protein showed constitutive kinase activity in the rice seedling leaf and stem. It was further identified that the 47-kDa protein kinase activity induced by cold in both the cytosolic and membrane fractions of the stem was strictly Ca²⁺-dependent. This CDPK activitiy increased in the presence of the Ca²⁺ ionophore A23187 in stem segments, whereas it was decreased by the Ca²⁺ channel blocker, LaCl₃, and the Ca²⁺ chelator, EGTA. The general protein kinase inhibitor, staurosporine, completely inhibited this CDPK activity in vitro, and both W7, a calmodulin antagonist, and H7, a protein kinase C inhibitor, could only partially decrease this activity. The protein phosphatase inhibitor, okadaic acid, increased CDPK activity. This CDPK activity was also induced by salt, drought stress and the phytohormone abscicic acid. Among the 18 rice varieties tested, this cold-induced 47-kDa CDPK activity was stronger in the cold-tolerant varieties than in the sensitive ones. Received: 13 August 1999 / Accepted: 24 January 2000     

水稻花药绒毡层及乌氏体的超微结构观察

在花粉母细胞期,水稻花药绒毡层细胞原生质浓,细胞器丰富,各轴向壁厚度较一致．随着药室腔扩大,绒毡层细胞体积迅速增大,且外切向壁增厚,径切向壁部分区域消失,细胞间形成原生质桥．在单胞花粉早期,乌氏前体排列于绒毡层内切向细胞膜内,随后移向膜外,且外侧增厚形成乌氏体．在花粉单核靠边期,绒毡层细胞的细胞器开始解体,到花粉充实期完全解体,但乌氏体结构直到花粉成熟保持不变．     

Desiccation Tolerance and Potential Longevity of Developing Seeds of Rice (Oryza sativa L.)

The longevity and desiccation tolerance of samples of seedsof a japonica rice (Oryza sativa L.) harvested serially duringdevelopment from plants grown in two temperature regimes, viz28/20 °C and 32/24 °C (12/12 h) were determined. Massmaturity (defined as the end of the seed-filling phase) occurred19·7 and 18·3 d after 50% anthesis, respectively.Longevity (determined at 40 °C with 15% moisture contentand quantified by the value of the constant K_i of the seed viabilityequation) improved during seed development and maturation until17 and 14 d after mass maturity in the cooler and warmer regimes,respectively, but declined thereafter. Changes in K_i with timewere similar in the two environments until mass maturity, butthe increase in K_i values after mass maturity was much greaterin the cooler regime. Tolerance of desiccation to low (4%) moisturecontents improved until 22 and 14 d after mass maturity in thecooler and warmer regimes, respectively, when maturation dryinghad reduced seed moisture contents naturally to 24 and 32% moisturecontent, respectively. Further delays to seed harvest reduceddesiccation tolerance, particularly in the warmer environment.Comparison among 15 samples of seeds harvested at differenttimes in the two environments showed a strong correlation (r= 0·947, P < 0·01) between longevity (K_i) anddesiccation tolerance (to 4% moisture content). Hence, it issuggested that the regulation of desiccation tolerance to lowmoisture contents and potential air-dry longevity during seeddevelopment and maturation determined here may have a commoncause.Copyright 1994, 1999 Academic Press Oryza sativa L., rice, desiccation tolerance, genebanks, seed development, seed longevity, temperature