雄性不育高粱（3197A）的热激蛋白与育性的关系

在我们以前研究热激(40℃)诱导雄性不育高粱3197A呈现雄性可育态时的内部物质的变化,即热激蛋白的表现的基础上,进一步研究观察到,在逐渐升温过程中,雄性不育高粱幼苗出现了热激蛋白的先后合成与消失的现象,即有阶段性变化的3次协同调控现象。雄性不育高粱与雄性可育高粱比较,在常温(28℃)不育系的自显影区带33KD、35KD比可育高粱的对应带强,在逐渐升温过程中,两个高粱系的这些蛋白带的差别逐渐缩小,到40℃时,两系的标记带图型趋于一致,表现了化学物质的变化与热激使不育系转变为可育态时的温度是一致的。比较高粱的总蛋白与可溶性蛋白,探明了某些蛋白为结合蛋白。用基因表达抑制剂探明热激反应可能与细胞核基因转录和转译两级调控有关。而与细胞质关系不大。试验探明逐渐升温到46—49℃温度区间,不育系幼穗的蛋白电泳图谱中出现特异的80KD热激蛋白,可育的保持系没有,表明此蛋白质与雄性不育有关。     

高粱热激蛋白(HSPs)的电泳分析与雄性不育性

本文以高粱雄性不育系为材料进行了热激蛋白的研究。苗期单向电泳表明,保持系在热激(40℃)过程中可溶性蛋白有24条带,对照(28℃)22条,出现2条带差异,不育系在热激后有33条带,比对照28条多5条,且有4条加强带,共9条带产生差异。苗期双向电泳,不育系在热激后比对照有19点产生差异,保持系热激后与对照比较有6点产生差异。表明双向电泳揭示了热激后蛋白质在分子量和电性方面有差异。花粉母细胞期幼穗的热激蛋白,不育系对照可溶性蛋白仅有3条带,热激后有11条,8条带有差异。保持系对照有10条带,热激后有15条,1条加强,共6条带有差异。幼穗期不育系蛋白质突出的缺少19kd以上的大分子量的带。从个体发育看,不育系由苗期可溶性蛋白比保持系带数多,发育到花粉母细胞期比保持系突出地减少,尤其是大分子量的蛋白带消失,热激后明显地增加了与可育的保持系相同的蛋白成份,表明不育系有其特殊的基因调控。     

高粱雄性不育系热激前后线粒体的变化与育性的关系

高粱雄性不育系3197A（3A）在花粉母细胞期进行热激处理后由不育变为可育。提取其处理小穗线粒体用流式细胞仪计数进行分析。结果发现经热激处理后,3A不育系的线粒体数目由1.55×106个／mg鲜穗增加至7.1×106／mg。同时对热激处理后3A的花粉育性及同工酶进行了研究,发现热激后3A的花粉粒变的饱满并可被I2朘I 溶液染色,其细胞色素氧化酶和过氧化物酶的电泳酶谱中出现与保持系3B一致的酶带。     

高梁雄性不育系热激前后线粒体的变化与育性的关系

高梁雄性不育系3197A(3A)在花粉母细胞期进行热激处理后由不育变为可育。提取其处理小穗线粒体用流式细胞仪计数进行分析。结果发现经热激处理后,3A不育系的线粒体数目由1.55×106个/mg鲜穗增加至7.1×106/mg。同时对热激处理后3A的花粉育性及同工酶进行了研究,发现热激后3A的花粉粒变的饱满并可被I2-KI溶液染色,其细胞色素氧化酶和过氧化物酶的电泳酶谱中出现与保持系3B一致的酶带。     

高粱3197A雄性不育系热激处理后代的可育性遗传

对高梁细胞质雄性不育系3197A经热激处理后获得的可育结实后代进行了连续多代自交及回交观察,结果表明,由热激诱导的可育性变化不能稳定遗传。从H2代开始,其可育率逐代降低。到H5代,分离出的可育株数仅占观察植株总数的1．9％。H5代可育株与不育株花药总RNA与HSP70热激蛋白基因探针Northern杂交结果表明,H3A可育株均能与HSP70基因探针杂交,显示较强的杂交信号。而其不育株,有的完全不能与HSP70基因探针杂交,有的显示微弱的杂交信号。说明3197A雄性不育系热激可育后代的遗传稳定性与HSP70基因的转录状况有关。     

热休克对于高粱细胞质雄性不育系线粒体数目及其育性的影响

对高粱雄性不育系3197A（3A）及其保持系3197B（3B）均分别进行热激处理,以常温处理为对照,以处于花粉母细胞期的小穗材料提取其线粒体,首次采用流式细胞仪测量线粒体的数量。首次观察到：经热激处理后,不育系3A线粒体的数目增加约3．5倍;相反,保持系3B在常温时,线粒体数量为3A的3．55倍,热激后却无明显变化（Fig.1)。同时取热激处理及常温下3A及3B的花药淀粉粒作碘化钾染色,结果表明：热激后3A花药中出现饱满淀粉粒（Fig.2);对其细胞色素氧化酶和过氧物酶同功酶的研究表明,热激后3A电泳酶谱中有新条带出现,而且与3B的带型一致（Fig.3)。     

抗阿维菌素朱砂叶螨的热激反应及热激蛋白

选用朱砂叶螨Tetranychus cinnabarinus阿维菌素抗性品系和敏感品系,测定了热预刺激后其在极限高温下的存活率,并应用SDS-PAGE技术研究了热激蛋白(HSPs)的种类及其含量。结果表明：非致死的热预刺激能显著提高朱砂叶螨耐极限温度的能力。两个品系在不同温度热激处理后,其蛋白质种类和含量发生了变化。正常情况下,朱砂叶螨敏感品系与阿维菌素抗性品系相比缺失8条条带;敏感品系热激后,增加了分子量分别为97.2,74.3,62.4,53.0和30.3 kDa的5条条带; 抗性品系热激后没有特异蛋白带的产生,但进一步高温胁迫后有些蛋白表达增强。此结果有助于解释朱砂叶螨抗性品系存在高温适合度优势现象。     

几种热激蛋白在细胞凋亡信号通路中的调控作用

热激蛋白(heat shock proteins, HSPs)作为进化保守的蛋白家族 之一,普遍存在于各种生物体中,并在生物体内发挥着重要的生理功能.大 量的实验证据表明,热激蛋白与细胞凋亡密切相关,参与细胞凋亡信号通 路的多个环节. 近年来有关该领域的研究已获得了重要的突破与进展.一方 面,热激蛋白主要起着抑制细胞凋亡、促进细胞存活的作用;另一方面, 某些热激蛋白又能够作为凋亡蛋白的分子伴侣,促进细胞凋亡,比如HSP70 能够激活DNase来促使细胞凋亡,线粒体内HSP60能够促进caspase依赖的细 胞凋亡途径.本文在阐明细胞凋亡信号通路的基础上,综述了近年来几种不 同热激蛋白家族（HSP90、 HSP70 、HSP60和小分子HSPs）在细胞凋亡调控 中作用的研究进展,重点阐述了几种主要热激蛋白与细胞凋亡信号通路上 相关因子的相互作用,并绘制了热激蛋白在细胞凋亡信号通路中的调控图 ,为进一步完善细胞凋亡调控网络研究提供一定的参考.     

雄性不育高粱线粒体与细胞核对热激的反应及其与育性关系的研究

由于我们发现热激(40℃)能诱导雄性不育高粱转变为雄性可育型的现象,故进一步探讨此现象的机理,研究了在热激条件下其细胞质(取其细胞质重要遗传物质——线粒体)中及细胞核中蛋白质的变化。试验结果,发现不育系的花粉母细胞期幼穗,在热激时细胞核中出现特异的80KD热激蛋白,且能被利福霉素、氯霉素所抑制。保持系在热激时细胞核中未出现此蛋白。80KD蛋白在结合部位存在的性质与其存在于细胞核中和线粒体中的现象是一致的。这种差异说明与雄性不育有关,且其原因是由细胞核与细胞质共同作用所致。     

热激处理与水稻雄性不育系育性转变关系的初步研究

本实验对水稻雄性不育系珍汕97A及其保持系在幼苗(2叶或3叶)和孕穗期(抽穗前8天)分别进行一次热激处理,并比较了其同工酶组成、花粉育性以及自交结实性的变化。同工酶分析结果表明,热激处理能使不育系与保持系花药的POX和Est的表达发生不同的改变,导致两系在这两种同工酶组成上的原有差异缩小;育性观察结果显示,在正常温度条件下,热激处理对珍汕97A的育性表达几乎无影响,而在有高温作用(穗分化发育阶段的日平均温度高于30℃)的条件下,3叶期幼苗往后的热激处理似乎能不同程度地提高珍汕97A的育性稳定性。简要讨论了热激处理与不育系育性转变间的关系。     

HSP_(70)基因表达对高粱育性的影响(英文)

高粱细胞质雄性不育系３１９７Ａ（３Ａ）在常温条件下是不育的（Ｆｉｇｓ．１１＆２）,经热激（４５℃）诱导不同程度地恢复了育性（Ｆｉｇｓ．１３＆４）,为研究其不育机理提供了线索。热激２ｈ后,３Ａ中即可产生一类线粒体热激蛋白（ＨＳＰｓ）。其中,分子量为７０ｋＤ的ＨＳＰ７０含量最高,也最为稳定。不过,３Ａ中ＨＳＰｓ的稳定性弱于保持系３１９７Ｂ（３Ｂ）（Ｆｉｇ．２,Ｐａｎｅｌｓ１～４）。放线菌素Ｄ抑制ＨＳＰｓ的合成,而氯霉素无此作用（Ｆｉｇ．２,Ｐａｎｅｌｓ５＆６）,表明：ＨＳＰｓ是由核基因编码、在细胞质中合成、再跨膜转运到线粒体中的。３Ａ幼穗经热激后,线粒体的总蛋白量猛增了２．７倍（Ｆｉｇ．３）,达到３Ｂ的水平,育性亦变为可育的。Ｆｉｇ．４表明：ＨＳＰ７０反义链ｃＤＮＡ（Ｒ１）能进入到３Ｂ花药细胞中,并与靶ＲＮＡ（ＨＳＣ７０ｍＲＮＡ）结合,而对照、正义链ｃＤＮＡ（Ｄ）链无此反应。由此、再增加一个通用保守序列的反义链ｃＤＮＡ（Ｒ２）、共两个探针（Ｒ１、Ｒ２）,可以检测到：３Ａ在常温下没有能力合成ＨＳＣ７０ｍＲＮＡ（Ｆｉｇ．５）,而在热激条件下,转变为有能力（Ｆｉｇ．６）。启示：３Ａ在热激条件下由不育转变为可育     

Developmental regulation of heat shock protein synthesis and HSP 70 RNA accumulation during postimplantation rat embryogenesis

Exposure of postimplantation rat embryos on days 9, 10, 11, and 12 of gestation to an in vitro heat shock of 43 degrees C for 30 min results in the induction of heat shock proteins (HSPs) in day 9 and 10 embryos, a severely attenuated response in day 11 embryos, and no detectable response in day 12 embryos. The heat shock response in day 9 embryos (presomite stage) is characterized by the synthesis of HSPs with molecular weights of 28-78 kDa. In heat shocked day 10 embryos, two additional HSPs are induced (34 and 82 kDa). In addition, two HSPs present on day 9 are absent on day 10. In day 11 heat shocked embryos, only three HSPs (31, 39, and 69 kDa) are induced, while in day 12 embryos no detectable HSPs are induced. Northern blot analysis of HSP 70 RNA levels indicates that the accumulation of this RNA, but not actin RNA, varies depending on developmental stage at the time of exposure to heat as well as the duration of the heat shock. Day 9 embryos exhibit the most pronounced accumulation of HSP 70 RNA while embryos on days 10-12 exhibit an increasingly attenuated accumulation of HSP 70 RNA, particularly after the more acute exposures (43 degrees C for 30 or 60 min). Thus, the ability to synthesize HSP 70 and to accumulate HSP 70 RNA changes dramatically as rat embryos develop from day 9 to day 12 (presomite to 31-35 somite stages).     

Possible cytoskeletal association of 69,000- and 68,000-dalton heat shock proteins and structural relations among heat shock proteins in murine mastocytoma cells

When murine mastocytoma cells (FMA 1) were heat shocked (42 degrees C for 4 h), nine heat shock proteins (HSPs) were detected by two-dimensional gel electrophoresis. Their apparent molecular weights were 100, 85, 69, 68, 32, 30, and 23 kDa (3 of 23 kDa). The structural homology of 4, 69, 68, 32, and 30 kDa, was demonstrated by two-dimensional tryptic peptide mapping. The 69- and 68-kDa HSPs were purified and rabbit antisera against these HSPs were prepared. A small fraction (less than 10%) of the 69- and 68-kDa HSPs were copurified with the microtubules and were present in the Triton X-100/KCl cytoskeletal fraction as shown by immunoblotting with the antiserum and by peptide mapping. Our results are consistent with the hypothesis of a cytoskeletal role for HSPs.     

玉米胚发育过程中热激蛋白的合成

35S-Met标记玉米胚蛋白合成结果表明,热激处理(42℃)与对照(25℃)的蛋白合成趋势相近,热激抑制16 DAP的蛋白合成,增加22和34 DAP蛋白合成.SDS-PAGE自显影图谱表明,热激诱导16DAP的胚合成86.4、80.0、73.2 kD等3种分子量较高的热激蛋白,22DAP后热激诱导合成86.4、80.0、73.2、24.4、18.2、16.8和13.6 kD等7种分子量的热激蛋白.2D-PAGE自显影图谱进一步显示,热激诱导22和28 DAP的胚合成近20种热激蛋白,其中超过10种为小分子热激蛋白.特异热激蛋白BiP(HsP70)、PDI(HsP60)Western blot表明,这2种热激蛋白在玉米胚发育过程均有高水平的表达,热激对其合成影响不明显.     

Heat and sodium arsenite act synergistically on the induction of heat shock gene expression in Xenopus laevis A6 cells

Heat shock protein (HSP) synthesis was studied in the Xenopus epithelial cell line A6 in response to heat and sodium arsenite, either singly or together. Temperatures of 33-35 degrees C consistently brought about the synthesis of HSPs at 87, 73, 70, 54, 31, and 30 kilodaltons (kDa), whereas sodium arsenite at 25-100 microM induced the synthesis of HSPs at 73 and 70 kDa. In cultures exposed to 10 microM sodium arsenite at 30 degrees C, HSP synthesis in the 68- to 73-kDa and 29- to 31-kDa regions was much greater than the HSP synthesis in response to each treatment individually. RNA dot blot analysis using homologous genomic subclones revealed that heat shock induced the accumulation of HSP 70 and 30 mRNAs. The sizes of the HSP 70 and 30 mRNAs determined by Northern hybridization were 2.7 and 1.5 kilobases, respectively. Sodium arsenite (10-100 microM) also induced the accumulation of both HSP 70 and 30 mRNAs. Finally, a mild heat shock (30 degrees C) plus a low concentration of sodium arsenite (10 microM) acted synergistically on HSP 70 and 30 mRNA accumulation in A6 cells. Thus sodium arsenite and heat act synergistically at the level of both HSP synthesis and HSP mRNA accumulation.     

Early and late heat shock proteins in wheats and other cereal species

Coleoptiles and roots of 3-day-old seedlings from five cereal species (Triticum aestivum L., T. durum Desf., Hordeum vulgare L., Secale cereale L., and Triticale) respond to heat shock at 40°C by synthesizing a new set of 13 strong bands (as revealed by one-dimensional sodium dodecyl sulfate gel electrophoresis) as well as some 20°C proteins. Heat shock proteins (HSPs) belong to three different size groups: high molecular mass HSPs in the 103 to 70 kilodalton range, intermediate molecular mass HSPs in the 62 to 32 kilodalton range, and low molecular mass HSPs about 17 to 16 kilodalton in size. At the beginning of the heat shock coleoptiles show a reduced ability to synthesize intermediate molecular mass HSPs but after 4 hours at 40°C they exhibit fully developed HSP patterns identical to that found in roots. Synthesis of early HSPs declines after 7 hours of treatment followed by the appearance of a new set of 12 protein bands (late HSPs) in the ranges 99 to 83, 69 to 35, and 15 to 14 kilodaltons. After 12 hours at 40°C, three other late HSPs of 89, 45, and 38 kilodalton are induced. The induction of late HSPs after 7 hours at 40°C appears to be associated with an enhancement of radioactive methionine incorporation into proteins.     

Expression of heat shock and cold shock proteins in the gorgonian Leptogorgia virgulata

In this study, we analyzed the response of the temperate, shallow-water gorgonian, Leptogorgia virgulata, to temperature stress. Proteins were pulse labeled with (35)S-methionine/cysteine for 1 h to 2 h at 22 degrees C (control), or 38 degrees C, or for 4 h at 12.5 degrees C. Heat shock induced synthesis of unique proteins of 112, 89, and 74 kDa, with 102, 98 and 56 kDa proteins present in the control as well. Cold shock from 22 degrees C-12.5 degrees C induced the synthesis of a 25 kDa protein, with a 44 kDa protein present in the control as well. Control samples expressed unique proteins of 38, and 33 kDa. Non-radioactive proteins expressed under the same conditions as above, as well as natural field conditions, were tested for reactivity with antibodies to heat shock proteins (HSPs). HSP60 was the major protein found in L. virgulata. Although HSP47, HSP60, and HSP104 were present in all samples, the expression of HSP60 was enhanced in heat stressed colonies, while HSP47 and HSP104 expression were greatest in cold shocked samples. Inducible HSP70 was expressed in cold-shocked, heat-shocked, and field samples. Constitutively expressed HSP70 was absent from all samples. The expression of HSP90 was limited to heat shocked colonies. The expression of both HSP70 and HSP104 suggests that the organism may also develop a stress tolerance response.     

Heat shock proteins and chilling sensitivity of mung bean hypocotyls

Excised mung bean (Vigna radiata L.) hypocotyl sections wereexposed to 40 C for up to 4 h in the presence or absence of50 µM cycloheximide (CHX) before being held at a non-chilling(20 C) or chilling (2.5 C) temperature. Mung bean hypocotyltissue is chilling sensitive, and the rate of solute leakageis highly correlated with the extent of chilling injury. A 3h heat shock at 40 C reduced chilling-induced solute leakageby up to 40%, but leakage was similar to non-heat-shocked hypocotylswhen CHX was present. Specific proteins were labelled when hypocotylswere exposed to [³⁵S] methionine during the last hour of heatshock. The nine most intense bands on the autoradiographs ofSDS-PAGE gels of extracted protein corresponded to molecularweights of 114, 79, 73, 70, 60, 56, 51, 46, and 18 kDa. The18 kDa band reached a maximum after 1 h at 40 C and then rapidlydecreased in intensity as the heat shock continued, becomingundetectable at 4 h. The four most intense bands after 3 h at40 C corresponded to molecular weights of 79, 70, 51, and 46kDa. The synthesis of these four hsps was markedly reduced whenthe hypocotyl sections were exposed to CHX during heat shock.During chilling for 6 d, the levels of hsps 79 and 70 remainedsignificantly higher in tissue that was heat shocked prior tochilling than in tissue that was not heat shocked. In contrast,the levels of hsps 51 and 46 were similar in bothheat-shockedand control tissues. Heat-shock-induced chilling tolerance waslost between 6 and 9 d ofstorage at 2.5 C; this loss coincidedwith the decay of hsps 79 and 70 to control levels. These resultssuggest that heat shock induces an increase in both chillingtolerance and the de novo synthesis of specific heat shock proteins;namely hsps 79 and 70. This is the first report showing a relationshipbetween heat-shock-induced chilling tolerance and specific heat-shock-inducedproteins. Key words: Ion leakage, protein synthesis, Vigna radiata     

Intracellular Distribution of Heat Shock Proteins in Pigeon Pea (Cajanus cajan)

The proteins synthesized In response to higher temperature In pigeon pea (Cajanus cajan) plants have been studied with respect to their Intracellular localization using root tissue. The heat shock proteins (hsps) of 18, 20, 22 and 24 kD were found to be associated with mitochondrial and membrane fractions, while the 60, 70 and 81 kD hsps were found In the soluble fraction. No evidence for the presence of hsps among the proteins synthesized in organello by isolated mitochondria could be obtained. Low molecular weight hsps (18, 20, 22 and 24 kD) were found associated with mitochondria Isolated from the heat shocked tissue suggesting that these hsps may have been transported post-translationally into mitochondria.     

Heat shock gene expression in Xenopus laevis A6 cells in response to heat shock and sodium arsenite treatments

Continuous exposure of a Xenopus laevis kidney epithelial cell line, A6, to either heat shock (33 degrees C) or sodium arsenite (50 microM) resulted in transient but markedly different temporal patterns of heat-shock protein (HSP) synthesis and HSP 70 and 30 mRNA accumulation. Heat-shock-induced synthesis of HSPs was detectable within 1 h and reached maximum levels by 2-3 h. While sodium arsenite induced the synthesis of some HSPs within 1 h, maximal HSP synthesis did not occur until 12 h. The pattern of HSP 70 and 30 mRNA accumulation was similar to the response observed at the protein level. During recovery from heat shock, a coordinate decline in HSPs and HSP 70 and 30 mRNA was observed. During recovery from sodium arsenite, a similar phenomenon occurred during the initial stages. However, after 6 h of recovery, HSP 70 mRNA levels persisted in contrast to the declining HSP 30 mRNA levels. Two-dimensional polyacrylamide gel electrophoresis revealed the presence of 5 HSPs in the HSP 70 family, of which two were constitutive, and 16 different stress-inducible proteins in the HSP 30 family. In conclusion, heat shock and sodium arsenite induce a similar set of HSPs but maximum synthesis of the HSP is temporally separated by 12-24 h.