钙调素mRNA和蛋白在水稻花药和雌蕊发育过程中的原位定位

利用ＲＮＡ原位杂交和免疫组织化学定位技术分别检测了钙调素ｍＲＮＡ和钙调素蛋白在水稻（ＯｒｙｚａｓａｔｉｖａＬ．）花药和雌蕊发育过程中的时空分布特征。钙调素基因在绒毡层、柱头、花粉管生长途径、退化助细胞以及维管薄壁细胞中大量表达,也可在小孢子母细胞、小孢子、花粉、反足细胞、卵细胞以及中央细胞中检测到。钙调素基因的表达强度随不同的发育阶段而变化：花药发育早期表达强,以后逐渐减弱并向特定部位集中,如绒毡层和花粉萌发孔等。胚胎发育早期,钙调素基因在胚乳细胞中的表达比原胚中强,而后期则在分化胚中比胚乳细胞中强。推测在有性生殖过程中,钙调素可能通过Ｃａ２＋ＣａＭ信号途径调节小孢子发育、花粉萌发、花粉管生长、受精以及物质运输等生理过程     

白芷培养细胞外钙调素结合蛋白的胶体金电镜定位研究

利用胶体金标主民技术制备了具有生物活性的植物Ｃａｍ－ＢＡＳ－ｇｏｌｄ探针,并用此探针建立了白芷愈伤组织培养细胞下调素结合蛋白的透射电镜标 记方法。标记结果显示,在１ｍｍｏｌ／ＬＣａ＾２＋存在下,用ＥＧＴＡ洗涤过的白芷愈伤组织培养细胞壁表面有金颗粒分布,而分别在含有ＥＧＴＡ、ＴＦＰ、过量未标记的ＣａＭ、ＣａＭ抗体存在的民政部下,用金示ＣａＭ探针进行标记,以及用的各对照组,细胞壁表面金颗粒则消失,说明     

水稻受精前后胚囊内钙调素分布的变化：免疫金电镜观察

用胶体金免疫电镜技术观察了水稻 (Oryzasativasubsp .japonica)受精前后胚囊内钙调素的分布变化。授粉后 ,卵细胞、助细胞和中央细胞内的钙调素较授粉前均有所增加。中央细胞内钙调素的增加要比卵细胞中约早 2h ,退化助细胞与宿存助细胞之间的钙调素含量无明显差异。授粉到受精期间 ,钙调素的主要分布形式由分散的单颗粒转变为聚集颗粒 ,受精完成后再变为分散的单颗粒形式。胚囊壁及珠心细胞的细胞壁和胞间隙中也观察到钙调素的分布和数量变化。初步讨论了胞内和胞外钙调素在水稻受精与合子形成中的作用。     

水稻受精前后胚囊内钙调素分布的变化:免疫金电镜观察

用胶体金免疫电镜技术观察了水稻(Oryza sativa subsp. japonica)受精前后胚囊内钙调素的分布变化.授粉后,卵细胞、助细胞和中央细胞内的钙调素较授粉前均有所增加.中央细胞内钙调素的增加要比卵细胞中约早2 h,退化助细胞与宿存助细胞之间的钙调素含量无明显差异.授粉到受精期间,钙调素的主要分布形式由分散的单颗粒转变为聚集颗粒,受精完成后再变为分散的单颗粒形式.胚囊壁及珠心细胞的细胞壁和胞间隙中也观察到钙调素的分布和数量变化.初步讨论了胞内和胞外钙调素在水稻受精与合子形成中的作用.     

白芷培养细胞外钙调素结合蛋白的胶体金电镜定位研究

利用胶体金标记技术制备了具有生物活性的植物CaM-BSA-gold探针,并用此探针建立了白芷愈伤组织培养细胞胞外钙调素结合蛋白(CaMBPs)的透射电镜标记方法。标记结果显示,在1mmol/L Ca~(2 )存在下,用EGTA洗涤过的白芷愈伤组织培养细胞壁表面有金颗粒分布,而分别在含有EGTA、TFP、过量未标记的CaM、CaM抗体存在的情况下,用金标CaM探针进行标记.以及用金标羊抗兔抗体替代金标CaM探针进行标记的各对照组,细胞壁表面金颗粒则消失,说明白芷愈伤组织培养细胞壁表面存在着CaM结合位点或CaMBPs。     

光敏核不育水稻花药IAA的免疫组织化学分析

首次应用免疫组织化学分析方法对长日和短日处理后的农垦５８Ｓ和对照农垦５８（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．ｓｕｂｓｐ．ｊａｐｏｎｉｃａ）花药中的ＩＡＡ进行了定位研究和相对水平的比较。结果表明,此方法可反映游离态ＩＡＡ在花药中的分布及其相对水平的变化。从雌雄蕊原基形成期至单核晚期的５个时期中,经长日照处理的农垦５８Ｓ花药中的ＩＡＡ水平都低于短日照处理的农垦５８Ｓ及在不同光周期处理下的农垦５８花药。对花     

玉米根尖细胞内钙调素的胶体金免疫电镜定位

钙调素(Calmodulin,简称CaM)抗体是研究CaM的特异性生物探针,常被用来研究CaM的定量和分布.CaM分布的研究不仅对揭示细胞内CaM的作用位点有重要意义,而且还可深入探讨CaM的功能.     

蚕豆保卫细胞中钙调素的免疫电镜定位

以蚕豆横切和平切气孔为材料,对钙调素进行了免疫胶体金电镜定位的结果表明:在蚕豆保卫细胞的细胞核、细胞质、细胞膜、叶绿体、液泡、高尔基体、细胞壁中都有金颗粒分布,在线粒体上的分布较少.     

胶体金免疫层析技术的应用与展望

本文主要介绍了胶体金免疫层析技术的发展及其在人体激素或疾病有关蛋白的检测、病原微生物有关蛋白或抗原抗体的检测、寄生虫相关抗原抗体的检测、药物浓度的监测等方面的情况,并对该技术作了展望。     

钙调素激酶在玉米体内的免疫组织化学定位

应用免疫组织化学定位方法研究了玉米体内钙调素激酶(CaM kinase,CaMK)的表达模式。结果表明CaMK广泛分布于玉米体内,但表达水平存在着明显时空差异。在营养器官中钙调素激酶主要分布于叶的维管束鞘细胞、侧根原基和根尖等部位,而其它部位没有检测到明显的分布。在生殖器官中,有大量钙调素激酶分布于幼胚及花药小孢子母细胞、四分体及绒毡层细胞中;在成熟胚囊的卵细胞、中央细胞以及二者的分界面上也有少量分布。这些结果为进一步探索钙调素激酶在植物体内的生理功能提供了重要线索。     

Cellular organisation and differentiation of organelles in pre-meiotic rice anthers

Pre-meiotic cellular organisation of rice anthers has a great significance in pollen formation. We have used a combination of confocal laser and transmission electron microscopy (TEM) to characterise and differentiate organelles in pre-meiotic rice anthers. Along with the characteristic organelles in the cytoplasm the epidermal cells of the pre-meiotic rice anther are coated on their outer surface by a conspicuous bi-lamellate cuticle. Chloroplasts of the endothecium contain immature grana, thylakoids and also starch granules. These plastids clearly contain photosynthetic pigments as shown by autofluorescence in confocal microscope studies. Both confocal and TEM studies reveal clusters of mitochondria in the middle layer. The tapetum contains electron opaque ribosomes, bundles of mitochondria and plastids. The nuclei of the tapetum occupy a large volume of the cytoplasm indicating the onset of mitotic prophase. Intense Rhodamine 123 staining reveals that a major portion of the structurally indistinguishable organelles that were seen throughout the densely ribosomic cytoplasm of sporogenous cells are mitochondria.     

Proteome analysis of male gametophyte development in rice anthers

We used proteomic analysis to investigate the changing patterns of protein synthesis during pollen development in anthers from rice plants grown under strictly controlled growth conditions. Cytological analysis and external growth measurements such as anther length, auricle distances and days before flowering were used to determine pollen developmental stages. This allowed the collection of synchronous anther materials representing six discrete pollen developmental stages. Proteins were extracted from the anther samples and separated by two-dimensional gel electrophoresis to produce proteome maps. The anther proteome maps of different developmental stages were compared and 150 protein spots, which were changed consistently during development, were analysed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry to produce peptide mass fingerprint (PMF) data. Database searches using these PMF data revealed the identities of 40 of the protein spots analyzed. These 40 proteins represent 33 unique gene products. Four protein spots that could not be identified by PMF analysis were analysed by N-terminal microsequencing. Multiple charge-isoforms of vacuolar acid invertase, fructokinase, beta-expansin and profilin were identified. These proteins are closely associated with sugar metabolism, cell elongation and cell expansion, all of which are cell activities that are essential to pollen germination. The existence of multiple isoforms of the same proteins suggests that during the process of pollen development some kind of post-translational modification of these proteins occurs.     

Genetic analysis of leaffolder resistance in rice

A double haploid(DH)population,which consists of 120 lines derived from anther culture of a typical indica and japonica hybrid'CJ06'/'TN1',was used to investigate the genetic basis for rice leaffolder resistance.Using a constructed molecular linkage map,five QTLs for rolled leaves were detected on chromosomes 1,2,3,4,and 8.The positive alleles from C J06 on chromosomes 3,4,and 8 in-creased the resistance to dee leaffolder,and the alleles from TN1 on chromosomes 1 and 2 also enhanced resistance to leaffolder.The interactions between QTLs were identified and tested,and four conditional interactions were acquired for resistance to rice leaffolder.These loci were located on chromosomes 2,9,10,and 11,respectively.QTL pyramiding indicated that the positive alleles affect resis-tance to leaffolder.The prospective application of this data in rice breeding was also discussed.     

水稻Ds插入淡绿叶突变体的鉴定和遗传分析

Ac/Ds插入突变是水稻基因功能鉴定的有力工具之一。文章从水稻中花11 Ds-T-DNA转化纯合体与Ac-T-DNA 转化纯合体的杂交群体中筛选到一个淡绿叶突变体。该突变体在三叶期由绿苗转为淡绿叶苗, 自然光照下突变体迅速焦枯, 但是在弱光照条件下, 突变体能缓慢生长至开花结实; 突变体光合作用特性研究表明该突变是典型的光抑制突变体。遗传分析表明该突变为Ds插入导致的隐性突变。     

Genetic analysis of leaffolder resistance in rice

A double haploid(DH)population,which consists of 120 lines derived from anther culture of a typical indica and japonica hybrid 'CJ06'/'TN1',was used to investigate the genetic basis for rice leaffolder resistance.Using a constructed molecular linkage map,five QTLs for rolled leaves were detected on chromosomes 1,2,3,4,and 8.The positive alleles from CJ06 on chromosomes 3,4,and 8 in-creased the resistance to rice leaffolder,and the alleles from TN1 on chromosomes 1 and 2 also enhanced resistance to leaffolde...     

Effect of iron plaque outside roots on nutrient uptake by rice (Oryza sativa L.). Zinc uptake by Fe-deficient rice

This solution culture study examined the effect of the deposition of iron plaque on zinc uptake by Fe-deficient rice plants. Different amounts of iron plaque were induced by adding Fe(OH)₃ at 0, 10, 20, 30, and 50 mg Fe/L in the nutrient solution. After 24 h of growth, the amount of iron plaque was correlated positively with the Fe(OH)₃ addition to the nutrient solution. Increasing iron plaque up to 12.1 g/kg root dry weight increased zinc concentration in shoots by 42% compared to that at 0.16 g/kg root dry weight. Increasing the amount of iron plaque further decreased zinc concentration. When the amounts of iron plaque reached 24.9 g/kg root dry weight, zinc concentration in shoots was lower than that in shoots without iron plaque, implying that the plaque became a barrier for zinc uptake. While rice plants were pre-cultured in –Fe and +Fe nutrient solution in order to produce the Fe-deficient and Fe-sufficient plants and then Fe(OH)₃ was added at 20, 30, and 50 mg Fe/L in nutrient solution, zinc concentrations in shoots of Fe-deficient plants were 54, 48, and 43 mg/kg, respectively, in contrast to 32, 35, and 40 mg/kg zinc in shoots of Fe-sufficient rice plants. Furthermore, Fe(OH)₃ addition at 20 mg Fe/L and increasing zinc concentration from 0.065 to 0.65 mg Zn/L in nutrient solution increased zinc uptake more in Fe-deficient plants than in Fe-sufficient plant. The results suggested that root exudates of Fe-deficient plants, especially phytosiderophores, could enhance zinc uptake by rice plants with iron plaque up to a particular amount of Fe.     

Variations in green plant regeneration response from anthers of indica rice and their hybrids with japonica cv. Taipei 309

The green plant regeneration ability from anthers of BR-7, a high yielding indica cultivar, Binnatoa (BA), a salt tolerant indica land race and IR-43 was tested in N6, M8, He2 and R2 media. The response was calculated on the basis of number of anthers producing green plants. The number of green plants per responding anther was also recorded. The response of BR-7 and BA was poor compared to the indica cultivar IR-43 in three of the media that were tested. In N6 medium, green plant regeneration of BA and BR-7 was respectively 10-fold and 100-fold less than the japonica cultivar Taipei 309 (T-309). No anther-derived green regenerant was obtained from another salt tolerant indica land race, Rajashail (RAJ). The N6 medium was selected to test green plant regeneration frequency from anthers obtained from the F1 crosses of T-309 × BR-7, T-309 × BA, T-309 × RAJ and T-309 × BR-7 AC regenerants backcrossed with BA. Our objective was to combine the salt tolerant trait of BA and the high yield of BR-7 in a single line. The intermediate crossing step with T-309 was performed to increase the green plant regenerability of the anthers. All F1 progeny from the crosses with T-309 showed significantly increased callus induction compared to the indica parent although the values were lower than the midparent means. Green plant regeneration compared to their respective indica parents either increased or decreased but never approached the level of T-309. This revised version was published online in June 2006 with corrections to the Cover Date.     

水稻糙米重金属Cd含量的QTL分析

镉(Cd²⁺)是一种分布较广泛、毒性较强的一种重金属, 文章利用韭菜青×IR26杂交衍生的一个重组自交系群体(Recombinant inbred lines, RIL)及构建的SSR分子遗传图谱, 对控制糙米中Cd²⁺含量的QTL进行分析, 为选育籽粒中Cd²⁺低吸收或低积累的水稻品种提供参考。结果表明, 在Cd²⁺胁迫(5 mg/kg)处理条件下, 共检测到2个与糙米Cd²⁺含量有关的QTLs, 分别位于水稻第11染色体上的标记RM6288-RM6544和RM167-RM5704之间, 其中qCCBR-11a对表型贡献率为11.17%, 加性效应0.089; qCCBR-11b对表型变异贡献率为7.66%, 加性效应0.075。相关分析显示, 糙米Cd²⁺含量与株高、每穗总粒数、每穗实粒数、结实率和千粒重等产量性状的相关性均不显著, 糙米中Cd²⁺含量是一个相对独立、由基因控制的遗传性状。