An efflux transporter of silicon in rice

Silicon is an important nutrient for the optimal growth and sustainable production of rice. Rice accumulates up to 10% silicon in the shoot, and this high accumulation is required to protect the plant from multiple abiotic and biotic stresses. A gene, Lsi1, that encodes a silicon influx transporter has been identified in rice. Here we describe a previously uncharacterized gene, low silicon rice 2 (Lsi2), which has no similarity to Lsi1. This gene is constitutively expressed in the roots. The protein encoded by this gene is localized, like Lsi1, on the plasma membrane of cells in both the exodermis and the endodermis, but in contrast to Lsi1, which is localized on the distal side, Lsi2 is localized on the proximal side of the same cells. Expression of Lsi2 in Xenopus oocytes did not result in influx transport activity for silicon, but preloading of the oocytes with silicon resulted in a release of silicon, indicating that Lsi2 is a silicon efflux transporter. The identification of this silicon transporter revealed a unique mechanism of nutrient transport in plants: having an influx transporter on one side and an efflux transporter on the other side of the cell to permit the effective transcellular transport of the nutrients.     

高等植物硅素营养研究进展

硅在地壳中含量为27.6%,大部分以二氧化硅（SiO2）的形态存在,能被植物吸收的有效硅只有50～250mg/kg,植物吸收硅后土壤溶液中硅的浓度仍能保持在一恒定水平.硅在植物内主要以二氧化硅胶（SiO2＊nH2O）的形态存在于表皮细胞和细胞壁上,含量随物种不同而差异很大,同时还受植株部位、栽培方式及环境条件等因素影响.植物吸收硅主要以单硅酸形式,不同植物吸收硅的能力不同,水稻具有主动吸收硅的能力,大多数植物被动吸收硅.硅素对植物的形态结构和体内其它元素产生影响,能调节植物的光合作用和蒸腾作用,能增强植物抗倒性和抗旱性,高浓度的硅对真菌孢子的萌发和菌丝的生长有抑制作用,硅素能够提高植物对病虫害的抗性,降低病虫害的发生.     

Biogenic nanostructured silica

Silicon is by far the most abundant element in the earth crust and also is an essential element for higher plants, yet its biology and mechanisms in plant tolerance of biotic and abiotic stresses are poorly understood. Based on the molecular mechanisms of the biosilicification in marine organisms such as diatoms and sponges, the cell wall template-mediated self-assembly of nanostructured silica in marine organisms and higher plants as well as the related organic molecules are discussed. Understanding of the templating and structure-directed effects of silicon-processing organic molecules not only offers the clue for synthesizing silicon-based materials, but also helps to recognize the anomaly of silicon in plant biology.     

2种非生物胁迫和7种激素对水稻OsAQP基因表达的影响

OsAQP是从cDNA文库中分离的一种新的水稻水通道蛋白基因,本实验室前期利用半定量RT-PCR技术发现该基因的表达与非生物胁迫、植物激素有关.本文采用qRT-PCR方法,检测了干旱、高盐胁迫以及ABA等7种植物激素对OsAQP在幼苗期水稻根和叶中表达的影响,结果显示干旱、高盐以及7种植物激素MeJA,SA,6-BA,GA,IAA,ABA,BR均能不同程度诱导OsAQP基因上调表达,在根中的诱导上调水平显著高于叶,提示该基因功能涉及生长发育、抗逆应答等多种生物学过程,且与水稻的根的关系更为密切.     

Overexpression of glutamine synthetases confers transgenic rice herbicide resistance

Glutamine synthetase (GS, E.C.6.3.1.2) is a key enzyme involved in the assimilation of inorganic nitrogen in higher plants and gram-negative microorganisms. GS is the targeting enzyme of a herbicide phosphinothricin (PPT) or Basta. In order to generate PPT-resistant transgenic rice via overexpression of GS, we constructed a plant expression vector p2GS harboring two different isoenzymes GS1 and GS2 cDNAs under the control of constitutive promoters of rice Act1 and maize Ubiquitin(Ubi) genes. The p2GS was introduced into rice genome by Agrobacterium-mediated transformation and confirmed by PCR and Southern blot hybridization. GS-transgene expression was first detected by Northern blot analyses. Results from Basta test indicated that GS-transgenic plants can tolerate as high as 0.3% Basta solution. In addition, our results also demonstrated that GS overexpression conferred transformed rice calli PPT resistance. Thus, GS cassette can serve as a selective marker gene instead of bar cassette for selection of PPT transformants.     

Obtaining transgenic rice resistant to rice fungal blast disease by controlled cell death strategy

The strategy of the two-component system,composed of Barnase and Barstar which encode RNase and a specific inhibitor to the RNase respectively, is adopted to obtain transgenic rice resistant to rice fungal blast disease. In this study, two chimeric promoters, induced by rice blast fungus pathogen (Magnaporthe grisea), are fused with Barnase respectively to construct two plant expression vectors, pWBNBS and pPBNBS together with the Barstar driven by CaMV 35S promoter. The resistance of the transgenic rice lines to rice blast fungus disease and rice blight disease are evaluated. The results show that (1) the expression of Barnase is induced in rice leaves when inoculated with the spores of Magnaporthe grisea; (2) the induced expression level of Barnase surpasses the level of Barstar, which elicits a similar hypersensitive response (HR) in the leaves, and the transgenic plant shows high resistance to the rice fungal blast disease; and (3) transgenic rice plants also show obvious resistance to rice bacterial blight disease. Taken together, these results suggest that the transgenic rice plants harboring this two-component system acquire relatively broad spectrum resistance against pathogens, especially high resistance to rice fungal pathogen.     

R gene expression induced by a type-III effector triggers disease resistance in rice

Disease resistance (R) genes in plants encode products that specifically recognise incompatible pathogens and trigger a cascade of events leading to disease resistance in the host plant. R-gene specificity is dictated by both host R genes and cognate avirulence (avr) genes in pathogens. However, the basis of gene-for-gene specificity is not well understood. Here, we report the cloning of the R gene Xa27 from rice and the cognate avr gene avrXa27 from Xanthomonas oryzae pv. oryzae. Resistant and susceptible alleles of Xa27 encode identical proteins. However, expression of only the resistant allele occurs when a rice plant is challenged by bacteria harbouring avrXa27, whose product is a nuclear localized type-III effector. Induction of Xa27 occurs only in the immediate vicinity of infected tissue, whereas ectopic expression of Xa27 resulted in resistance to otherwise compatible strains of the pathogen. Thus Xa27 specificity towards incompatible pathogens involves the differential expression of the R gene in the presence of the AvrXa27 effector.     

硅对叶螨危害后棉花防御酶活性的调控作用及其与抗虫性的关系

以棉花品种新陆早33号感染土耳其斯坦叶螨（Tetranychus turkestani（Ugarov ＆ Nikolski））作个案研究,阐明硅提高棉花抗螨性的生理机理。网室条件下采用微区试验设计,硅肥采用喷施和基施处理,研究不同时间棉花的虫害指数、干物质积累量和硅含量,以及与植物抗性密切相关的过氧化物酶（POD）、多酚氧化酶（PP0）和苯丙氨酸解氨酶（PAL）活性变化。结果显示：不同施硅处理缓解了棉花叶片受叶螨危害的症状,在接种叶螨第6天硅处理降低了叶片受害指数;施硅处理提高了PPO和PAL的活性,而降低了参与清除植物体内活性氧功能的POD活性。硅能够调控棉花与抗虫有关的酚类物质代谢过程,参与植物防卫反应而增强棉花对土耳其斯坦叶螨的抗性。     

Adaptation of photoperiodic control pathways produces short-day flowering in rice

The photoperiodic control of flowering is one of the important developmental processes of plants because it is directly related to successful reproduction. Although the molecular genetic analysis of Arabidopsis thaliana, a long-day (LD) plant, has provided models to explain the control of flowering time in this species, very little is known about its molecular mechanisms for short-day (SD) plants. Here we show how the photoperiodic control of flowering is regulated in rice, a SD plant. Overexpression of OsGI, an orthologue of the Arabidopsis GIGANTEA (GI) gene in transgenic rice, caused late flowering under both SD and LD conditions. Expression of the rice orthologue of the Arabidopsis CONSTANS (CO) gene was increased in the transgenic rice, whereas expression of the rice orthologue of FLOWERING LOCUS T (FT) was suppressed. Our results indicate that three key regulatory genes for the photoperiodic control of flowering are conserved between Arabidopsis, a LD plant, and rice, a SD plant, but regulation of the FT gene by CO was reversed, resulting in the suppression of flowering in rice under LD conditions.     

bar基因表达框和完整质粒转化对水稻农艺性状的变异效应比较

通过分析转基因植株的株高、分蘖数、结实率、千粒重四个主要农艺性状的变异,比较基因枪法转化的基因表达框(仅含启动子、基因开放阅读框和终止子序列)和完整质粒两种基因载体形式对水稻农艺性状的变异效应.结果表明,与非转基因亲本相比,转基因植株的千粒重和分蘖数与对照无显著差异;株高变异也不大,17个转基因株系中,仅基因表达框转化的2个株系XFb-36和XFb-63株高变矮;而结实率变异最大,有9个转基因株系结实率显著降低,变异率达50%以上.总体上看,基因表达框和完整质粒两种基因载体形式对水稻农艺性状的变异效应差异不明显,基因枪转化对水稻农艺性状的变异效应主要表现为降低植株的育性,转基因水稻植株的农艺性状变异主要来源于转基因过程中组织培养引起的无性系变异.     

水稻组蛋白单泛素化连接酶基因(OsHUB1)的克隆及其表达分析

对模式植物拟南芥的研究发现,AtHUB1基因参与调控植物对灰霉病的抗性.为了研究水稻的抗病机理,笔者利用同源搜索从水稻数据库中获得了目标基因的序列,并通过RT-PCR技术从水稻中克隆了组蛋白单泛素化连接酶基因,命名为OsHUB1.该片段包含1个2 655 bp的开放阅读框,编码了1个885氨基酸的多肽.与NCBI数据库的比对结果表明:OsHUB1的氨基酸序列与短柄草(Brachypodium distachyon)、葡萄(Vitis vinifera)、杨树(Populus tomentosa)、大豆(Glycine max)和拟南芥(Arabidopsis thaliana)的泛素E3连接酶的同源性分别为78%,68%,68%,67%和62%.这些不同来源的组蛋白单泛素化连接酶都含有一个高度保守的RING指结构域.半定量表达分析结果表明:OsHUB1基因能够被水杨酸(SA)和茉莉酸(JA)诱导表达,说明OsHUB1基因可能通过SA和JA介导的信号转导途径参与水稻的抗病反应.     

逆境胁迫下水稻蛋白质组学研究进展

水稻在生长发育过程中要面对各种非生物胁迫如干旱、高盐、温度、重金属和生物胁迫如病虫害等,通过观察水稻在各种胁迫条件下的蛋白质组表达情况,动态分析水稻的蛋白质组变化,对新逆境相关蛋白质进行分离与功能鉴定,这不仅能揭示参与胁迫耐受的蛋白质翻译后调控机制,而且可以增进对耐受胁迫分子机理的认识.综述了水稻应答逆境胁迫蛋白质组学研究的最新研究进展,探讨了存在的主要问题.     

Over-expression of an Arabi-dopsisd δ-OAT gene enhances salt and drought tolerance in transgenic rice

δ-OAT, ornithine-δ-aminotransferase, is the key enzyme involved in proline biosynthesis. In this study the Arabidopsis δ-OAT gene was transferred into rice (Oryza sativa L. ssp japonica cv. Zhongzuo 321), whose successful integration was demonstrated by PCR and Southern blot analysis. The over-expression of the gene in transgenic rice was also confirmed. Biochemical analysis showed that, under salt or drought stress conditions, proline contents in the leaves and roots in transgenic rice plants were 5- to 15-fold of those in non-transgenic controls. Under stress conditions, germinating rate of transgenic lines is higher than that of controls. Although the growth of rice plants tested were more and more retarded with the increasing of NaCI concentration, the transgenic plants grow faster compared to the controls under the same stress condition. Meanwhile, the resistance to KCl and MgSO4 stresses was also found enhanced in transgenie rice. Furthermore, the over-expression of δ-OAT also improved the yield of transgenic plants under stress conditions. The average yield per plant of transgenic lines increases about 12%--41% more than that of control line sunder 0.1 mol/L NaCI stress. These data indicated that the over-expression of δ-OAT, with the accumulation of proline, resulted in the enhancement of salt and drought tolerance and an increase of rice yield, which is of significance in agriculture.     

Production of bacterial blight resistant lines from somatic hybridization between Oryza sativa L. and Oryza meyeriana L

Novel bacterial blight (BB) resistance gene(s) for rice was (were) introduced into a cultivated japonica rice variety Oryza sativa (cv. 8411), via somatic hybridization using the wild rice Oryza meyeriana as the donor of the resistance gene(s). Twenty-nine progenies of somatically hybridized plants were obtained. Seven somatically hybridized plants and their parents were used for AFLP (amplified fragment length polymorphism) analysis using 8 primer pairs. Results confirmed that these plants were somatic hybrids containing the characteristic bands of both parents. The morphology of the regenerated rice showed characters of both O.sativa and O.meyeriana. Two somatic hybrids showed highest BB resistance and the other 8 plants showed moderate resistance. The new germplasms with highest resistance have been used in the rice breeding program for the improvement of bacterial blight resistance.     

唐山市水稻田土壤施硅肥对作物产量的影响

唐山市各县区长期种植水稻 ,逐渐表现出缺硅现象 :群体植株高度下降 ,茎叶变软 ,叶片披垂 ,易感病 ,籽粒空秕率高 ,导致产量降低。水稻田灌溉水有效硅含量 :沿海地区平均 1 2 92mg/kg ;内陆地区平均 2 37mg/kg。水稻田土壤有效硅含量 :沿海地区平均 74 57mg/kg ;内陆地区平均 59 63mg/kg。水稻田施用硅肥的最佳施肥量 :沿海地区施有效硅 1 2 1 5kg/hm2 ;内陆地区底施有效硅 1 61 6kg/hm2 。水稻田施硅增产的因子有 4个方面 :1、促进根系发育 ,增加白根数量 ,减少红、黑根数量 ,提高根系的吸收能力 ;2、提高植株有效分蘖和成穗率 ,减少养分消耗 ;3、增加植株高度 ,使茎叶健壮 ,提高抗病性 ;4、从产量结构分析 ,穗数、穗粒数、千粒重均增加 ,从而增高单位面积产量     

外源胁迫对水稻内源茉莉酸和水杨酸的影响

茉莉酸和水杨酸是植物防御反应中的重要分子，为研究茉莉酸和水杨酸在水稻防御反应中的作用，采用了色谱法测定了此两种激素在外源植物生长调节剂胁迫下的水平变化。我们采用的此方法简单，灵敏度高，再现性好，此两种激素含量变化可得到准确分离与测定。我们的结果表明在水稻中茉莉酸和水杨酸的作用可能不同与其它植物。首先，水杨酸含量变化较缓慢，其次，茉莉酸含量变化较明显。此结果表明水稻中茉莉酸和水杨酸的作用可能不同与烟草和拟南介，在信号传递过程中可能起到正面的调节作用。     

Transformation of GbSGT1 gene into banana by an Agrobacterium-mediated approach

SGT1 is a homologue of the yeast ubiquitin ligase-associated protein. It controls some protein degradation and activates defense pathway in plants. Cotton GbSGT1 gene (Gossypium barbadense) has been isolated and characterized in previous work. In this study, the plant expression vector pBSGT1 with bar gene as a selection agent was constructed and transgenic banana was obtained via Agrobacterium-mediated transformation with the assistance of particle bombardment and screened with PCR and Basta spreading on banana plant leaves. Estimating of transgenic banana plants for resistance to Panama wilt is in progress.