藻红蓝蛋白裂合异构酶E基因突变体的构建、表达与酶活性检测

通过诱变分别得到PecE的N端,C端和中间位置缺失突变的一系列突变基因的编码产物,测定了这5种缺失蛋白质的酶活性的大小。实验结果显示：来自层理鞭枝藻(M．laminosus,UTEX 1931)的PecE[简称PecE(UTEX 1931)]与来自层理鞭枝藻(M．laminosus,．PCC 7603)的PecE[简称：PecE(PCC 7603)]有81％的同源性。PecE(UTEX1931)／PeeF(PCC 7603)的酶催化活性与PecE(PCC 7603)／PecE(PCC 7603)的酶催化活性相当。PecE(UTEX 1931)的N端缺失22个氨基酸的蛋白质显示出了约20％的催化活性;而N端缺失36个氨基酸、56个氨基酸的蛋白质,C端缺失23个氨基酸的蛋白质和PecE(PCC 7603)中间缺失27个氨基酸的蛋白质均丧失活性,结果对酶的结构以及对酶活性的影响提供了重要信息。     

柞蚕核型多角体病毒(ApNPV)转移载体质粒pAp M2614的组建

自从美国科学家G.Smith等首次建立苜蓿尺蠖核型多角体病毒(AcNPV)转移载体表达系统以来,已被广泛用于外源基因的表达,成为世界上一新的具有巨大潜力的载体表达系统。为了进一步提高表达产量,降低成本,日本科学家前田进建立了家蚕核型多角体病毒(BmNPV)载体表达系统,并获得了高效表达。柞蚕是我国特产,以蛹滞育越冬,保存时间长,个体大,可工厂化生产。因此,组建柞蚕NPV转移载体,进而建立该载体表达系统,是目前利用昆虫活体为宿主进行外源基因表达较理想的昆虫杆状病毒载体表达系统。     

大豆根系质外体铁库的累积及其在缺铁时被利用的规律

采用营养液培养法研究了在不同程度的缺铁条件下,大豆（Ｇｌｙｃｉｎｅｍａｘ（Ｌ．）Ｍｅｒｒ．）根系质外体铁库累积与利用的规律,及其在缓解植物缺铁胁迫方面的作用。结果表明,缺铁处理下,大豆根系质外体铁库不断被再利用直至枯竭,根系还原力与过氧经物酶活性呈升降有序的周期性变化,从而一定程度上缓解缺铁胁迫,延缓植株缺铁症状的出现,新叶叶绿素和活性铁含量缓慢下降,而低铁条件下（供应１０＾－６ｍｏｌ／ＬＦｅＥＣ     

苏云金芽胞杆菌HD-73菌株sigL基因突变体的特性分析

[目的]通过分析苏云金芽胞杆菌sigL基因突变体的特征,进一步明确sigL基因在苏云金芽胞杆菌中的功能.[方法]测定了苏云金芽胞杆菌HD-73菌株,sigL基因缺失突变菌株和互补菌株在不同营养成分的培养基中的生长曲线以及在不同氮源条件下的生长情况.分别将调控aco操纵子(编码3-羟基丁酮脱氢酶系统)的转录调节基因acoR和调控bkd操纵子(编码催化支链脂肪酸合成的酶系统)的转录调节基因bkdR的启动子与lacZ基因融合,并转入出发菌株和sigL突变体中,测定β-半乳糖苷酶的活性.[结果]sigL突变体不能利用精氨酸、脯氨酸、缬氨酸、异亮氨酸、谷氨酰胺、苯丙氨酸、蛋氨酸、色氨酸为唯一的氮源;β-半乳糖苷酶的活性分析表明:在sigL突变体中acoR基因和bkdR基因的启动子活性降低.序列比对分析表明:Bt中的AcoR和BkdR的蛋白结构域与依赖于σL的转录调节因子的保守序列相似.[结论]苏云金芽胞杆菌中sigL基因的缺失可能阻碍了某些重要碳、氮源参与的代谢途径.在Bt中AcoR和BkdR是依赖于σL的转录调节因子.     

水稻根系铁膜形成及对磷吸收的影响

水稻(Oryzasativa)是全球重要的粮食作物。合理施肥是保障水稻持续稳产的必要农艺措施。磷是水稻生长必需的营养元素之一,主要通过水稻根系吸收。而水稻长期生长在淹水环境中,其根系表层形成富含铁氧化物的胶膜,并在水稻根际磷的迁移和转化过程中扮演关键角色。该文综述了生物和非生物因素对水稻铁膜形成和转化的影响,探讨了铁膜对磷吸收及转运的影响,并对后续研究提出建议,旨在为阐明水稻根际铁磷互作机制提供理论依据。     

不同生态修复方式对土壤团聚体稳定性及胶结物质的影响

土壤团聚体作为土壤结构的基本单元,其结构稳定性对边坡生态修复的效果与可持续性均有重要影响,但胶结物质在边坡生态修复中的作用尚未完全明确,为探明生态修复边坡土壤团聚体稳定性、胶结物质特征及二者之间的关系,进而为评估生态修复效果提供科学依据。选择宜昌市8种不同类型的生态修复边坡进行了实验,分析土壤团聚体稳定性和分形维数及胶结物质的含量,并分析各胶结物质对团聚体稳定性的影响。结果表明,(1)生态修复显著地改善了团聚体的粒径分布及稳定性特征,平均重量直径>0.25mm团聚体分量和分形维数三个指标均显著优于裸露边坡。(2)整体上生态修复显著提升了各粒径团聚体内的有机碳含量,且大团聚体内的有机碳含量要高于微团聚体,而铁铝氧化物主要向细粒径团聚体富集(3)冗余分析结果显示铁铝键结合态有机碳、钙结合态有机碳、有机碳是影响生态修复边坡土壤团聚体粒径分布及稳定性的关键因素。研究可为后续的生态修复工程提供理论支持和技术指导。     

铲形杰氏涡虫(单肠目,达氏科)新纪录的形态学描述及其分子系统学分析

本文对分布于广东省深圳市侨城湿地(113°58'E,22°31'N)咸淡水的一种单肠目Rhabdocoel涡虫进行了形态、发育及18S r DNA与28S r DNA分子系统学的分析与比较。结果表明,此涡虫为达氏科Dalyelliidae铲形杰氏涡虫Gieysztoria knipovici Beklemischev,1953,为中国新纪录种。该物种模式标本分布于欧洲黑海近岸。本研究详细描述了其外部形态与内部结构特征,首次发现该涡虫精巢超长,前端达咽部,后端至腹侧,呈长条形(该属涡虫2个精巢呈球形或椭圆形,位于肠后腹侧);除了骨质阴茎之外还具有可再生的骨质阴茎鞘。阴茎鞘位于骨质阴茎外侧,半包裹骨质阴茎,呈长条形骨质结构。交配后的个体常在交配囊内发现阴茎鞘1~5个。鉴于该物种特殊的形态学特征,其分类地位还有待进一步研究。     

控释尿素施用方式及用量对夏玉米氮肥效率和产量的影响

在大田条件下以不施氮为对照,研究了施氮量为75和150 kg N·hm^-2条件下,普通尿素底施、控释尿素底施和侧施对夏玉米光合性能、氮肥效率和产量的影响.结果表明：与普通尿素相比,相同施氮水平下,包膜控释尿素处理玉米果穗叶光合速率、叶绿素含量、硝酸还原酶活性、籽粒灌浆速率均显著提高,玉米籽粒产量比普通尿素平均提高9.5%;与控释尿素侧施相比,底施使玉米籽粒产量提高6.2%.包膜控释尿素处理的氮肥农学利用率（AE）和偏生产力（PFP）比普通尿素平均分别提高74.5%和11.0%;与包膜控释尿素侧施相比,底施使AE和PFP分别提高26.8%和5.7%.控释尿素施用量较高时,玉米光合性能得到改善,产量、AE和PFP显著提高;与控释尿素侧施相比,底施对玉米产量、AE和PFP的增加效果更显著.     

Phenotypic characterization of sym 21, a gene conditioning shoot-controlled inhibition of nodulation in Pisum sativum cv. Sparkle

E132 ( sym 21) is a stable pleiotropic mutant of Pisum sativum cv. Sparkle obtained by mutagenesis with ethyl methane sulfonic acid. The line forms few nodules and short, highly branched roots. Microscopy studies revealed that infection by rhizobia is normal, and low nodulation is mainly due to a low rate of emergence of the nodule meristems. E132 shoots depressed nodulation on Sparkle stocks, whereas in reciprocal grafts more nodules formed on E132 stocks than on control roots or self-grafted Sparkle plants. Nodule number on the mutant was slightly increased by exogenous ethylene inhibitors, which, however, did not alter the root phenotype.     

Molecular analysis of a null mutant for pea (Pisum sativum L.) seed lipoxygenase-2

A mutant line of Pisum fulvum was identified that lacked seed lipoxygenase-2 (LOX-2). The mutant phenotype was introgressed into a standard Pisum sativum cv. Birte to provide near-isogenic lines with or without seed LOX-2. Genetic analyses showed the mutation to behave as a single, recessive Mendelian gene. Northern and dot-blot analyses showed a large reduction in LOX-2 mRNA from developing seeds of the LOX-2-null mutant. A restriction fragment length polymorphism associated with the 5 end of the LOX-2 gene(s) co-segregated with the null phenotype, indicating that the reduction of LOX-2 mRNA was neither a consequence of deletion of the LOX genes nor a consequence of the action of a genetically distant regulatory gene. Analysis of the 5-flanking sequences of LOX-2 genes from Birte and the near-isogenic LOX-2-null mutant revealed a number of insertions, deletions and substitutions within the promoter from the LOX-2-null mutant that could be responsible for the null phenotype. Incubation of crude seed LOX preparations from Birte and the LOX-2-null mutant showed that the latter generated relatively less 13-hydroperoxides and also produced relatively more hydroxy- and ketoacid compounds that have implications for the fresh-frozen pea industry.     

褐藻胶寡糖对豌豆种子萌发和幼苗的某些生理特性的影响

褐藻胶寡糖(ADO)对豌豆种子萌发和幼苗生长有促进作用,种子发芽率升高,幼苗高度.根长、生物量均有增加;幼苗叶中叶绿素含量增高;叶片净光合速率、胞间CO_2浓度、气孔导度和蒸腾速率增加;根系活力增强;种子中α和β-淀粉酶活性增强。低浓度ADO的效果明显优于高浓度的ADO,其中以0.125?O的效果最佳。     

A chemically induced new pea (Pisum sativum) mutant SGECdt with increased tolerance to, and accumulation of, cadmium

BACKGROUND AND AIMS: To date, there are no crop mutants described in the literature that display both Cd accumulation and tolerance. In the present study a unique pea (Pisum sativum) mutant SGECd(t) with increased Cd tolerance and accumulation was isolated and characterized. METHODS: Ethylmethane sulfonate mutagenesis of the pea line SGE was used to obtain the mutant. Screening for Cd-tolerant seedlings in the M2 generation was performed using hydroponics in the presence of 6 microm CdCl2. Hybridological analysis was used to identify the inheritance of the mutant phenotype. Several physiological and biochemical characteristics of SGECd(t) were studied in hydroponic experiments in the presence of 3 microm CdCl2, and elemental analysis was conducted. KEY RESULTS: The mutant SGECd(t) was characterized as having a monogenic inheritance and a recessive phenotype. It showed increased Cd concentrations in roots and shoots but no obvious morphological defects, demonstrating its capability to cope well with increased Cd levels in its tissues. The enhanced Cd accumulation in the mutant was accompanied by maintenance of homeostasis of shoot Ca, Mg, Zn and Mn contents, and root Ca and Mg contents. Through the application of La(+3) and the exclusion of Ca from the nutrient solution, maintenance of nutrient homeostasis in Cd-stressed SGECd(t) was shown to contribute to the increased Cd tolerance. Control plants of the mutant (i.e. no Cd treatment) had elevated concentrations of glutathione (GSH) in the roots. Through measurements of chitinase and guaiacol-dependent peroxidase activities, as well as proline and non-protein thiol (NPT) levels, it was shown that there were lower levels of Cd stress both in roots and shoots of SGECd(t). Accumulation of phytochelatins [(PCcalculated) = (NPT)-(GSH)] could be excluded as a cause of the increased Cd tolerance in the mutant. CONCLUSIONS: The SGECd(t) mutant represents a novel and unique model to study adaptation of plants to toxic heavy metal concentrations.     

Gibberellic acid effects on the ultrastructure of endocarp cells of unpollinated ovaries of Pisum sativum

Ultrastructural changes of endocarp cells of unpollinated pea ( Pisum sativum L. cv. Alaska) ovaries treated and non-treated with gibberellic acid (GA₃) were studied. Following treatment with GA₃, the differentiation of cells of the middle zone of the endocarp into sclerenchyma occurred in 3 phases: initial, elongation and maturation. In the initial phase a proliferation of endocarp cells was produced in both GA₃treated and non-treated ovaries. Changes in the cell membrane system (Golgi apparatus, endoplasmic reticulum, plasmalemma) and the formation of primary and secondary cell wall were observed in the 2nd and 3rd phases of treated ovaries. In nontreated ovaries senescence followed the initial phase. These results indicate that GA₃ induced the processes involved in the elongation and maturation phases.     

Expression of gibberellin mutations in fruits of Pisum sativum L.

The gibberellin (GA) economy of young pea (Pisum sativum L.) fruits was investigated using a range of mutants with altered GA biosynthesis or deactivation. The synthesis mutation lh-2 substantially reduced the content of both GA₄ and GA₁ in young seeds. Among the other synthesis mutations, ls-1, le-1 and le-3, the largest reduction in seed GA₁ content was only 1.7-fold (le-1), while GA₄ was not reduced in these mutants, and in fact accumulated in some experiments (compared with the wild type). Mutation sln appeared to block the step GA₂₀ to GA₂₉ in young pods and seeds, but not as strongly as in older seeds. Mutations ls-1, le-1 and le-3 markedly reduced pod GA₁ levels, but pod elongation was not affected. After feeds of [¹³C,³H]GA₂₀ to leaves, the pods contained ¹³C,³H-labelled GA₂₀, GA₁, GA₂₉ and GA₈₁, and the seeds, [¹³C,³H]GA₂₀ and [¹³C,³H]GA₂₉. These findings are discussed in relation to recent suggestions regarding the role and origin of GA₁ in pea fruits. Received: 6 June 1997 / Accepted: 15 July 1997     

A relationship between irradiation, carbohydrates and rooting in cuttings of Pisum sativum

Rooting ability was studied for cuttings derived from pea plants ( Pisum sativum , L. cv. Alaska) grown in controlled environment rooms. When the cuttings were rooted at 70 μmol m⁻² s⁻, ¹ (photosynthetic photon flux density) or more, a stock plant irradiance at 100 μmol m⁻² s⁻¹ decreased rooting ability in cuttings compared to 5 μmol m⁻², s⁻¹, However, cuttings rooted at 160 μmol m⁻² s⁻¹ formed more roots compared to 5 (μmol m⁻² s⁻¹. Although a high irradiance increased the number of roots formed, it could not overcome a decreased potential for root formation in stock plants grown at high irradiance. Light compensation point and dark respiration of cuttings decreased by 70% during the rooting period, and the final levels were strongly influenced by the irradiance to the cuttings. Respiratory O₂ uptake decreased in the apex and the base of the cutting from day 2 onwards, whereas a constant level was found in the leaves. Only the content of extractable fructose, glucose, sucrose and starch varied during the early part of the rooting period. We conclude that the observed changes in the cuttings are initiated by excision of the root system, and are not involved in the initiation of adventitious roots.     

Gene expression during two alternative pathways of ovary development in Pisum sativum: fruit development and ovary senescence

Pea ovaries are induced to enter a fruit development pathway involving physiological and morphological changes by pollination or application of plant growth regulators. In the absence of these stimuli, overies stop growing and enter an alternative pathway of senesecence that leads to their degeneration. We have used two dimensional polyacrylamide gel electrophoresis in search of molecular changes underlying fruit development and ovary senescence at the level of total accumulated proteins, newly synthesized proteins, and translatable, RNA populations. We have found changes in gene expression during the processes of ovary formation and ovary senescence. Stimuli that induce fruit set do not appreciably alter the overall patterns of synthesized proteins or translatable RNAs, indicating that fruit development is apparently a natural continuation of ovary formation. However, ovary senescence is an alternative pathway that involves the presence of new RNA messengers and proteins as well as the disappearance of others. These changes were detected earlier than any morphological or structural changes could be observed in the ovary.