Characterization and mapping of a spotted leaf mutant in rice (Oryza sativa)

Spotted leaf mutant belongs to a class of mutants that can produce necrotic lesions spontaneously in plants without any attack by pathogens. These mutants have no beneficial effect on plant productivity but provide a unique opportunity to study programmed cell death in plant defense responses. A novel rice spotted leaf mutant (spl30) was isolated through low-energy heavy ion irradiation. Lesion expression was sensitive to light and humidity. The spl30 mutant caused a decrease in chlorophyll and soluble protein content, with marked accumulation of reactive oxygen species (ROS) around the lesions. In addition, the spl30 mutant significantly enhanced resistance to rice bacterial blight (X. oryzae pv. oryzae) from China (C1–C7). The use of SSR markers showed that the spl30 gene was located between markers XSN2 and XSN4. The genetic distance between the spl30 gene and XSN2 and between spl30 and XSN4 was 1.7 cM and 0.2 cM, respectively. The spl30 gene is a new gene involved in lesion production and may be related to programmed cell death in rice. The ability of this mutant to confer broad resistance to bacterial blight provides a model for studying the interaction between plants and pathogenic bacteria.     

Proteome analysis and characterization of phenotypes of lesion mimic mutant spotted leaf 6 in rice

Rice spotted leaf 6 (spl6) mutant produces lesions caused by spontaneous cell death in the absence of pathogenic infection. Expression of this genetic trait was developmentally programmed. After the tillering stage, small red and brown lesions were initiated in groups on the leaf blade. Eventually, the lesions formed parallel lines along the midrib of the leaf. Under light and transmission electron microscopy, we observed that thylakoid membranes of mesophyll chloroplasts were progressively damaged in the nonspotted section of the mutant leaf. However, chloroplasts were absent in the mesophyll cells of the spotted area of the spl6 mutant. These results indicated that lesion formation of the spl6 mutant might be caused by oxidative burst. Proteome analysis revealed that 159 protein spots were up or downregulated in comparison between spotted leaves of the spl6 mutant plants and normal leaves of the wild type. Among them, protein disulfide isomerase (PDI), transketolase, thioredoxin peroxidase (TPX), ATP synthase, RuBisCO large subunit, and RuBisCO activase small subunit were not identified in the spl6 mutant but were abundant in the wild type. Especially, the absence of TPX and PDI might be the cause of the failure to protect cells against oxidative burst resulting in degradation of the thylakoid membranes and leading to programmed cell death and lesion development.     

Comparative Leaf Epidermal Anatomy of Mutants of Barley (Hordeum vulgare L. 'Himalaya') which Differ in Leaf Length

We wished to determine the nature of differences in epidermalcell numbers and dimensions between leaves of different lengthin mutants of barley (Hordeum vulgare L. ‘Himalaya’).Three comparisons were made: leaf one (L1)vs. leaf four (L4);wild typevs. nine dwarf mutants and wild typevs. a slender mutant.L1 was shorter than L4, and for most lines this was associatedwith a change in epidermal cell number for the blade, and inboth cell number and length for the sheath. Compared to wildtype, the smaller leaves of dwarf plants generally had shorterand fewer cells in both blade and sheath. The blade of slenderplants was the same length (L1) or longer (L4) than wild type,while the sheath was longer than that of wild type for bothL1 and L4. Slender plants had longer but fewer cells than thewild type along the blade of L1, and shorter but more cellsfor the blade of L4. In the sheath, slender plants had longerand more (L1) or fewer (L4) cells than did the wild type. ForL1, variation in blade width amongst the barley lines was associatedwith a change in file width and file number. For L4, blade widthvaried only with file number, except for slender plants wherenarrow blades were associated with reduced file width. Hencethere was no consistent correlation between changes in cellsize or cell (or file) number with changes in leaf length orwidth. Differences depended on the leaf (L1vs. L4), leaf part(bladevs. sheath), and the nature of the mutation (dwarfvs.slender). Barley (Hordeum vulgare L. ‘Himalaya’); leaf epidermis; dwarf mutant; slender mutant     

水稻斑点叶突变体spl101和spl102的筛选及候选基因鉴定

斑点叶突变体是指植物在正常生长条件下叶片或叶鞘上自发形成、且与病原菌侵染产生的病斑类似的一类突变体,筛选并研究斑点叶突变体对揭示植物抗病反应机理具有重要意义。为了进一步研究斑点叶的形成机制,本文通过EMS诱变品种宽叶粳(KYJ),筛选得到两个斑点叶突变体spl101和spl102。这两个突变体在生长发育晚期(抽穗期以后)形成严重的类病斑。遗传分析表明,spl101和spl102均受隐性单基因控制。利用Mutmap方法对候选基因进行克隆,结果显示,spl101和spl102的候选基因均为OsEDR1,该基因与类病斑发生有关。在spl101中,OsEDR1基因突变发生在第6外显子和第6内含子的交接处,该突变导致第6内含子的错误识别,最终造成移码突变。在spl102中,OsEDR1基因突变发生在第10外显子上,导致一个苯丙氨酸(F)变成半胖氨酸(C)。因此,本研究鉴定了两个新的OsEDR1等位突变,对OsEDR1抗病反应机理的进一步研究及丰富水稻种质资源具有积极意义。同时验证了利用Mutmap方法克隆水稻突变基因的有效性。     

铜绿假单胞菌对镉胁迫苗期水稻根系活力及叶片生理特性的影响

为揭示耐镉铜绿假单胞菌缓解镉胁迫水稻的生理效应,以无镉处理为对照,通过添加菌液、空载体、菌剂及20μmol·L^-1 Cd进行水培试验,分析了菌株对苗期水稻根系活力及叶片生理特性的影响.结果表明:镉胁迫显著抑制了水稻的根系活力,降低了叶片光合效率、抗氧化酶活性及可溶性蛋白、类黄酮与总酚含量,提高了叶片丙二醛(MDA)和超氧阴离子(O2-)含量.与镉处理相比,添加菌液、菌剂处理的水稻根系活力分别提高了36.1%~42.5%、49.4%~53.0%;叶片净光合速率提高了118.5%~147.1%、137.6%~156.9%;可溶性蛋白含量提高了37.0%~49.3%、37.7%~72.6%.菌剂处理的水稻叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性分别比Cd处理提高了36.9%~42.6%、82.7%~92.6%、43.3%~52.2%,菌液处理的SOD、POD、CAT则分别比Cd处理提高了25.8%~36.6%、40.9%~55.9%、24.0%~29.2%,菌剂对水稻叶片抗氧化酶的促进效应显著高于菌液;菌剂、菌液处理的水稻叶片MDA含量分别比Cd处理降低了44.8%~54.7%、29.4%~41.9%;O2-含量减少了9.9%~10.2%、3.0%~7.1%;菌剂处理后类黄酮、总酚含量分别比Cd处理提高了125.4%~135.7%、100.8%~119.4%;菌液处理后则分别提高了139.4%~146.7%、115.0%~134.7%.可见,铜绿假单胞菌及其菌剂通过提高苗期水稻根系活力、光合作用促进了苗期水稻的生长.铜绿假单胞菌通过增强水稻抗氧化酶活性、提高类黄酮和总酚等抗氧化物质含量,表现出显著的缓解镉胁迫效应.     

205国道两侧农田土壤和水稻叶片及糙米中重金属含量的空间分布特征

对205国道无林带典型路段东西两侧200 m范围内的农田表层土壤、稻(Oryza sativa L. )叶及糙米中Al、Cd、Cr、Cu、Fe、Ni、Pb、Zn和As的含量及空间分布规律进行了分析研究.结果表明,205国道两侧农田表层土壤中Cd、Cr、Cu、Ni、Pb和Zn的含量达到土壤环境质量一级标准(GB15618-1995),As含量超过土壤环境质量三级标准.西侧表层土壤中的Al、Cr、Fe、Ni、Zn和As含量,稻叶中的Al、Cd、Cr、Fe和Ni含量及糙米中的Al、Cd、Cr、Cu、Fe和Ni含量均比东侧高,表明路西侧受汽车尾气扩散沉降影响比东侧明显,并与该路段主风向为偏东风有关;西侧表层土壤中的Ni和Zn含量及东侧表层土壤中的Cd含量、西侧稻叶中的Al和Zn含量、西侧糙米中的Fe含量和东侧糙米中的Al含量均有随路侧距离增加而显著递减的规律,负相关关系显著或极显著;稻叶和糙米中某些重金属的含量有明显的峰值区域,均位于路侧10～20 m区域内,并与土壤中的含量有显著或极显著的正相关关系;在糙米中未检出As,且Al、Cd、Cr、Cu和Zn含量均未超出国家食品卫生标准,但部分样本Ni和Pb含量略有超标.主成分分析结果显示,稻叶中的Pb主要来源于土壤,而稻叶中其他重金属含量明显受到公路环境污染物扩散沉降影响.研究结果显示,重金属元素已在205国道两侧的农田表层土壤中显著累积,其中As的积累最严重;稻叶和糙米中重金属的含量水平明显受到公路汽车尾气扩散沉降的影响,且扩散沉降集中在距离公路边缘10～20 m的区域内.     

Molecular re-confirmation and floral characteristics of drooping leaf (DL) mutants generated by insertional mutagenesis in rice

Transposon tagging and insertional mutagenesis provide one of the most powerful tools in gene function studies. Here, we report a comparison between two novel drooping leaf (DL) mutants from transposon and T-DNA insertion lines of rice. DL is distinct from well-known ABC genes and a member of the YABBY gene family, and it is closely related to the CRABS CLAW (CRC) gene of Arabidopsis thaliana. Based on phenotypic analysis, DL regulated midrib formation by promoting cell proliferation in the central region of rice leaf and was necessary for the specification of carpel identity. We identified two DL mutants by screening the Ac/Ds and T-DNA insertional mutant pool of rice. Flanking sequence tag analysis indicated that both Ds and T-DNA segments were inserted in the promoter region at 3.4 kbs and 5.4 kb upstream, respectively, of the previously known OsYABBY domain. Interestingly, the progenies of DL lines of two different pools showed various degrees of leaf drooping and abnormal carpel formation. Flower structures revealed that there were more than two stigmas with normal stamens and pistils per panicle in the Ds-induced mutants. However, T-DNA induced mutant had extra stamens with staminoid carpels. These results indicate that the promoter region of DL plays an important function in regulating anther and carpel formation.     

开花期低温胁迫对水稻花粉性状及剑叶理化特性的影响

以耐冷水稻品种996和冷敏感品种4628为材料,开花期在人工气候室进行7 d低温（06:00-8:00和19:00-23:00,19 ℃;08:00-10:00和16:00-19:00,21 ℃;10:00-16:00,23 ℃;23:00-06:00,17 ℃）和适温（06:00-8:00和19:00-23:00,24 ℃;08:00-10:00和16:00-19:00,26 ℃;10:00-16:00,30 ℃;23:00-06:00,22 ℃)处理,研究了低温胁迫对水稻花药开裂、花粉性状及剑叶叶绿素、可溶性糖和可溶性蛋白质含量、膜透性等理化特性的影响.结果表明：开花期低温胁迫导致水稻花药开裂系数、花粉萌发率显著下降,中部和下部颖花的不育花粉率增加.996的花药开裂系数和花粉萌发率显著高于4628,表明耐冷品种996在开花期低温胁迫下能保持较好的花粉散落特性和花粉萌发特性.低温胁迫下,996的剑叶可溶性蛋白质和游离脯氨酸含量及其增幅显著高于4628,而丙二醛含量和相对电导率及其增幅却显著低于4628,说明耐冷品种在低温胁迫下的保护性反应更迅速和强烈,其膜结构及功能更稳定.     

Morphological and physiological characteristics of a root-hairless mutant in rice (Oryza sativa L.)

This paper reports morphological and physiological characteristics of a first root-hairless mutant (RH2) of rice (Oryza sativa L.), which can be useful in advancing knowledge on the role of root hairs in water and nutrient uptake, and genetics of root hairs. The mutant was selected among NaN₃ mutagenized progeny of the rice cultivar Oochikara. Microscopic observations showed absence of root hairs in RH2. At the seedling stage, RH2 showed shorter seedling height and shorter roots compared to the wild type variety Oochikara. Because of the differences in seedling growth, all comparisons between Oochikara and RH2 in uptake-related characters were made on the basis of values adjusted by the dry weight of either the shoot or the root. When grown at low water potential in soil, Oochikara and RH2 were similar in shoot water content and transpiration per unit shoot dry weight, and similarly, at low water potential in solution culture, there was no significant difference between Oochikara and RH2 in transpiration per unit shoot dry weight. These results suggest that at the seedling stage, root hairs do not significantly contribute to uptake of water. In solution culture, Oochikara and RH2 did not significantly differ in phosphate uptake per unit root dry weight. This result supports the previous work that root hairs do not contribute to phosphate uptake in solution culture. Regarding to response to plant hormones, RH2 showed a higher level of resistance to two synthetic auxins, 2,4-dichlorophenoxyacetic acid (2,4-D) and 1-naphthaleneacetic acid (NAA) than Oochikara. NAA treatment induced very short root hairs in RH2, suggesting that the absence of root hairs in RH2 may be due to a shortage of endogenous auxin. Genetic analysis showed that the root hairless character in RH2 is inherited as a single recessive gene.     

Developmental and physiological correlates of leaf size in Hyeronima alchorneoides (Euphorbiaceae)

The tropical emergent tree Hyeronima alchorneoides has large decreases in leaf size with tree age: 1200 cm(2) at 1 yr, 900 cm(2) at 3 yr, 200 cm(2) at 11 yr, and 80 cm(2) in old (>30 yr) individuals. We tracked leaf growth and physiological attributes on trees of three different ages (1, 3, and 11 yr) to determine the developmental basis and functional consequences of this variation. Leaves on young trees grew faster and sustained maximum rates of leaf expansion longer than leaves on older trees. Leaf mass per area (LMA) did not differ among age classes. Maximum photosynthetic rates reflected differences in leaf nitrogen concentration, in which leaves from the lower crown of younger trees outperformed those at a comparable crown position in older trees. One-year-old trees had the lowest stomatal conductance and the greatest instantaneous water use efficiency. Ontogenetic plasticity in mature leaf size, structure, and physiology may be a balance between the advantages conferred by rapid height growth when trees are young and the benefits derived from producing branches that increase light harvesting ability as trees reach the canopy.     

Comparative physiological and immunological characteristics of carps (cyprinidae) grown in aquaculture with different types of food

On the basis of the hematological, cytochemical, and biochemical blood indices, it has been established that the adaptive capabilities in common carp Cyprinus carpio and grass carp Ctenopharyngodon idella are higher than in silver carp hybrid Hypophthalmichthys molitrix × H. nobilis, while the potential for growth is, by contrast, higher in silver carp hybrid.     

水稻类病斑突变体的生理与遗传分析

从全基因组水平上筛选获得了10个籼稻和8个粳稻类病斑(lesion resembling disease,lrd27-44)突变体.从突变体性状受环境影响敏感程度可以分为环境钝感型和环境敏感型.从发育进程可以分为全生育期类病斑型,营养生长阶段起始类病斑型和生殖生长阶段起始类病斑型.病斑的光诱导表明病斑由受光信号激发的程序性细胞死亡引起,而不受损伤诱导.对其中4个突变体lrd32,lrd39,lrd40和lrd42的遗传分析结果表明,这些类病斑性状由1或2对隐性基因控制.两个突变体lrd37和lrd40表现出对白叶枯病菌的广谱抗病性,有关基因定位克隆正在进行中.     

湖南安息香属植物的叶片比较解剖学研究

利用光学显微镜和扫描电镜对湖南产安息香属10种植物叶表皮微形态和解剖结构进行了观察和测量。结果表明:(1)表皮均由单层细胞构成,细胞形状为多边形和无规则形,垂周壁式样为平直、弓形、波状;(2)气孔器大小及密度在种间的差异明显;(3)气孔外拱盖内缘主要为波状和浅波状,气孔器外围角质纹饰以越南安息香的鸟巢状和栓叶安息香的碗状为显著区别特征;(4)老鸹铃、垂珠花和白花龙的气孔器保卫细胞两极有"T"形加厚;(5)栓叶安息香和大果安息香叶上表皮由大型细胞组成;(6)叶肉分化为栅栏组织和海绵组织,二者的厚度及比值在种间有明显的差异,其中越南安息香栅栏组织分化为2层;(7)大果安息香和芬芳安息香的主脉维管组织构成一圈封闭的环状结构,而其他种则为开放的半圆形结构;(8)叶片及主脉的厚度在种间差异明显。安息香属植物的表皮微形态及解剖结构特征比较稳定,可作为种间鉴定和分类的重要依据。     

Expression of snowdrop lectin (GNA) in transgenic rice plants confers resistance to rice brown planthopper

Snowdrop lectin ( Galanthus nivalis agglutinin; GNA) has been shown previously to be toxic towards rice brown planthopper ( Nilaparvata lugens ; BPH) when administered in artificial diet. BPH feeds by phloem abstraction, and causes ‘hopper burn’, as well as being an important virus vector. To evaluate the potential of the gna gene to confer resistance towards BPH, transgenic rice ( Oryza sativa L.) plants were produced, containing the gna gene in constructs where its expression was driven by a phloem-specific promoter (from the rice sucrose synthase RSs1 gene) and by a constitutive promoter (from the maize ubiquitin ubi1 gene). PCR and Southern analyses on DNA from these plants confirmed their transgenic status, and that the transgenes were transmitted to progeny after self-fertilization. Western blot analyses revealed expression of GNA at levels of up to 2.0% of total protein in some of the transgenic plants. GNA expression driven by the RSs1 promoter was tissue-specific, as shown by immunohistochemical localization of the protein in the non-lignified vascular tissue of transgenic plants. Insect bioassays and feeding studies showed that GNA expressed in the transgenic rice plants decreased survival and overall fecundity (production of offspring) of the insects, retarded insect development, and had a deterrent effect on BPH feeding. gna is the first transgene to exhibit insecticidal activity towards sap-sucking insects in an important cereal crop plant.     

Effects of Angular Leaf Spot and Rust on Leaf Gas Exchange and Yield of Common Bean (Phaseolus Vulgaris)

Isolated and interactive effects of angular leaf spot (caused by Phaeoisariopsis griseola) and rust (caused by Uromyces appendiculatus) on leaf gas exchange and yield was studied in common bean (Phaseolus vulgaris L. cv. Carioca) plants. Gas exchange was measured on 37, 44, 51, and 58 d after planting using a portable photosynthesis system. The inoculation of plants with P. griseola (P), U. appendiculatus (U), and the combination of both pathogens (P+U) caused a significant reduction of net photosynthetic rate (P _N) and yield. The reduction of stomatal conductance (g _s), P _N, and yield was higher under P and combination of P+U than under U treatment. By effect of U, the reduction on yield was higher than the reductions on gas exchange parameters. On the treatment P+U, a reduction of 23 % in P _N and a correspondent reduction of 32 % in yield was observed. The interactive effects of the pathogens on yield could be explained in part by the decreases in g _s and in P _N of diseased bean leaves. The combined effect of both diseases on yield and gas exchange parameters suggests an antagonistic interaction.