β-Glucosidase treatment and infestation by the rice brown planthopper Nilaparvata lugens elicit similar signaling pathways in rice plants

β-Glucosidase has been reported to induce the production of herbivore-induced plant volatiles. However, how it works remains unclear. Here, we investigated the levels of salicylic acid （SA）, jasmonic acid （JA）, ethylene, and H2O2, all of which are known signaling molecules that play important roles in induced plant defense in rice plants treated with β-glucosidase, and compared these to levels in plants infested by the rice brown planthopper Nilaparvata lugens （Stal）. Results showed that wounding and treatment by β-glucosidase increased the levels of SA, ethylene, and H2O2, but not JA, in all plants compared to control plants. The signaling pathways activated by β-glucosidase treatment are similar to those activated by an infestation by N. lugens, although the magnitude and timing of the signals elicited by the two treatments are different. This may explain why both treatments have similar volatile profiles and are equally attractive to the parasitoid Anagrus nilaparvatae Pang et Wang.     

Salicylic acid and ethylene signaling pathways are involved in production of rice trypsin proteinase inhibitors induced by the leaf folder Cnaphalocrocis medinalis （Guenee）

The roles of signaling pathways in the production of trypsin proteinase inhibitors (TrypPIs) in rice infested by the leaf folder (LF) Cnaphalocrocis medinalis were studied. Infestation by LF increased TrypPI levels in the leaves of rice plants at the tillering, booting and flowering stages but decreased TrypPI levels at the ripening stage; TrypPI levels in rice stems did not increase at any developmental stage. Infestation by LF at the tillering stage systemically increased TrypPI levels in leaves but not in stems; it also enhanced salicylic acid (SA) levels in leaves and stems, and the ethylene level released from plants. However, LF infestation did not increase JA concentrations. Exogenous application of SA or ethylene enhanced TrypPI levels in the leaves and stems of plants at the tillering stage, whereas treatment with both SA and ethylene induced lower levels of TrypPIs than treatment with SA or ethylene alone, suggesting an antagonistic effect of SA and ethylene on TrypPIs induction. The results suggest that both SA and ethylene signaling pathways are involved in the production of TrypPIs in rice induced by LF; moreover, the antagonistic effect of SA and ethylene may explain the changes in TrypPI levels seen at different plant developmental stages and in different organs.     

Attractiveness of tobacco volatiles induced by <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> and <Emphasis Type="Italic">Helicoverpa assulta</Emphasis> to <Emphasis Type="Italic">Campoletis chlorideae</Emphasis>

The attraction of Helicoverpa armigera-and Helicoverpa assulta-induced and mechanical damage-induced tobacco volatiles to Campoletis chlorideae was investigated, and the induced volatiles were analyzed. In windtunnel, C. chlorideae was strongly attracted by herbivoreinduced tobacco volatiles. Mechanically damaged tobacco leaves, whether treated with caterpillar regurgitant or water,were more attractive to the parasitoid than undamaged tobacco leaves. GC-MS analysis revealed that only 4 compounds were released from undamaged tobacco leaves,whereas 13 compounds were commonly emitted from herbivore-infested and mechanically damaged tobacco leaves.Compound β-pinene was specifically induced by the infestation of H. armigera, and (Z)-3-hexenal was only induced by the infestation of H. armigera and H. assulta, whereas hexyl acetate was only induced by mechanical damage. Tobacco leaves infested by H. armigera and H. assulta released larger amounts of volatiles than undamaged tobacco leaves did.Tobacco leaves treated with artificial damage plus caterpillars regurgitant or water emitted the same levels of volatiles,which were higher than that emitted by undamaged tobacco leaves. The emission amounts of single compounds were also different between differently treated plants. The differences were large between herbivore-induced and mechanical damage-induced compounds, and small between H. armigeraand H.assulta-induced compounds, and among compounds emitted from mechanically damaged plants treated with water or caterpillar regurgitant.     

Caterpillar-induced nocturnal plant volatiles repel conspecific females

Plants respond to insect herbivory by synthesizing and releasing complex blends of volatile compounds, which provide important host-location cues for insects that are natural enemies of herbivores. The effects of these volatile blends on herbivore behaviour have been investigated to only a limited extent, in part because of the assumption that herbivore-induced volatile emissions occur mainly during the light phase of the photoperiod. Because many moths-whose larvae are some of the most important insect herbivores-are nocturnal, herbivore-induced plant volatiles have not hitherto been considered to be temporally available as host-location cues for ovipositing females. Here we present chemical and behavioural assays showing that tobacco plants (Nicotiana tabacum) release herbivore-induced volatiles during both night and day. Moreover, several volatile compounds are released exclusively at night and are highly repellent to female moths (Heliothis virescens). The demonstration that tobacco plants release temporally different volatile blends and that lepidopteran herbivores use induced plant signals released during the dark phase to choose sites for oviposition adds a new dimension to our understanding of the role of chemical cues in mediating tritrophic interactions.     

Involvement of Jasmonate-signaling pathway in the herbivore-induced rice plant defense

The expression patterns of eight defense- related genes in the herbivore-infested and jasmonate- treated (jasmonic acid, JA and its derivative MeJA) rice leaves were analyzed using RT-PCR. The results showed that Spodoptera litura Fabricius (Lepidoptera: Noctuidae) herbi-vory induced the expression of lipoxygenase (LOX) and al-lene oxide synthase (AOS) genes that are involved in the jasmonate-signaling pathway. Moreover, S. litura damage resulted in the expression of farnesyl pyrophosphate syn-thase (FPS), Bowman-birk proteinase inhibitor (BBPI), phenylalanine ammonia-lyase (PAL) and other rice defense- related genes that were also induced by aqueous JA treat-ment or gaseous MeJA treatment. These indicated that in rice leaves, the JA-related signaling pathway was involved in the S. litura-induced chemical defense. Mechanical damage and brown planthopper (BPH), Nilaparvata lugens (St錶) (Homoptera: Delphacidae) damage induced the expression of LOX gene, but both treatments did not induce the expression of AOS gene. However, BPH damage induced the expression of acidic pathogen-related protein 1 (PR-1a), Chitinase (PR-3), and PAL genes, which is involved in the salicylate- signaling pathway. It was suggested that salicylate-related signaling pathway or other pathways, rather than jas-monate-signaling pathway was involved in the BPH-induced rice plant defense.     

Effects of herbivore-induced rice volatiles on the host selection behavior of brown planthopper, Nilaparvata lugens

It has been suggested that herbivore would react to volatiles produced by herbivore infested plant due to potential change, either positive or negative, in the acceptability of the host plant. This hypothesis was tested for the brown planthopper (BPH) in the laboratory. Sixteen components of the headspace volatiles from rice seedlings with different treatments were collected with SPME and Tenax-TA trap and analyzed with GC and GC-MS. Significant differences in volatile emissions were observed for rice plants with different treatments. Undamaged control plants, mechanically damaged plants and the plants infested by BPH for 1 or 2 d emitted much lower amounts of volatiles compared to the plants infested by BPH for 3 or 5 d. The plants infested by BPH for 3 or 5 d emitted several volatiles that were not detected in undamaged control plants, mechanically damaged plants or the plants infested by BPH for 1 or 2 d. Spodoptera litura infested plants released much higher amounts of volatiles than those in all other treatments, and the contents of several green leaf volatiles, methyl salicylate and terpenoids increased dramatically. In dual-choice flight tunnel experiments, adult BPH females showed no significant preference between the untreated healthy plants and mechanically damaged plants or the plants infested by BPH adult females. However, rice plants damaged by S. litura had a clearly repellent effects on BPH adult females compared to healthy undamaged plants, mechanically damaged plants or the plants infested by BPH.     

Ethylene signaling in rice

Ethylene regulates many aspects of growth, development and responses to environmental stresses in plants. Its signaling pathway has been established in model dicotyledonous plant Arabidopsis. However, its roles and signal transduction in monocotyledous rice plant remain largely unknown. In this review, we summarize the current advances in rice ethylene signaling studies and compare these with the results from Arabidopsis and other plants. Most of the components homologous to those in Arabidopsis ethylene signaling pathway have been found in rice, including five ethylene receptors, OsEIN2, OsEIL1, and OsERFs. Rice ethylene receptors are functionally more divergent than that of Arabidopsis. OsEIN2 and OsEIL1 display limited roles in regulation of rice ethylene responses compared with their Arabidopsis orthologs. ERF-like proteins OsERF1 and OsEBP-89 appear to be involved in rice ethylene signaling. However, whether they are activated through OsEIN2 and OsEIL1-mediated pathway needs further studies. Given that rice uses ethylene to control many processes that do not exist in Arabidopsis, it seems that new components or new mechanisms may exist in rice ethylene signaling pathway.     

Effects of herbivore-induced rice volatiles on the host selection behavior of brown planthopper, Nilaparvata lugens

It has been suggested that herbivore would react to volatiles produced by herbivore infested plant due to potential change, either positive or negative, in the acceptability of the host plant. This hypothesis was tested for the brown planthopper (BPH) in the laboratory. Sixteen components of the headspace volatiles from rice seedlings with different treatments were collected with SPME and Tenax-TA trap and analyzed with GC and GC-MS. Significant differences in volatile emissions were observed for rice plants with different treatments. Undamaged control plants, mechanically damaged plants and the plants infested by BPH for 1 or 2 d emitted much lower amounts of volatiles compared to the plants infested by BPH for 3 or 5 d. The plants infested by BPH for 3 or 5 d emitted several volatiles that were not detected in undamaged control plants, mechanically damaged plants or the plants infested by BPH for 1 or 2 d. Spodoptera litura infested plants released much higher amounts of volatiles than those in all other treatments, and the contents of several green leaf volatiles, methyl salicylate and terpenoids increased dramatically. In dual-choice flight tunnel experiments, adult BPH females showed no significant preference between the untreated healthy plants and mechanically damaged plants or the plants infested by BPH adult females. However, rice plants damaged by S. litura had a clearly repellent effects on BPH adult females compared to healthy undamaged plants, mechanically damaged plants or the plants infested by BPH.     

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.     

Transgenic rice homozygous lines expressing GNA showed enhanced resistance to rice brown planthopper

Mature seed-derived calli from two elite Chinese japonica rice (Oryza sativa L.) cultivars Eyi 105 and Ewan 5 were co-transformed with two plasmids, pWRG1515 and pRSSGNA1, containing the selectable marker hygromycin phosphotransferase gene (hpt), the reporter β-glucuronidase gene (gusA) and the snowdrop (Galanthus nivalis) lectin gene (gna) via particle bombardment. 61 independent transgenic rice plants were regenerated from 329 bombarded calli. 79% transgenic plants contained all the three genes, revealed by PCR/Southern blot analysis. Western blot analysis revealed that 36 out of 48 gna-containing transgenic plants expressed GNA (75%) at various levels with the highest expression being approximately 0.5% of total soluble protein. Genetic analysis confirmed Mendelian segregation of transgenes in progeny. From the R2 generations whose R1 parent plants showing 3:1 Mendelian segregation patterns, we identified five independent homozygous lines containing and expressing all the three transgenes. Insect bioassay and feeding tests showed that these homozygous lines had significant inhibition to rice brown planthopper (Nilaparvata lugens, BPH) by decreasing BPH survival and overall fecundity, retarding BPH development and declining BPH feeding. These BPH-resistant lines have been incorporated into rice insect resistance breeding program. This is the first report that homozygous transgenic rice lines expressing GNA, developed by genetic transformation and through genetic analysis-based selection, conferred enhanced resistance to BPH, one of the most damaging insect pests in rice.     

三种水稻挥发物对稻田节肢动物群落和嗅觉行为的影响

茉莉酸甲酯、水杨酸甲酯和芳樟醇是水稻受虫害诱导后产生的重要挥发物性物质。它们通过直接或间接的途径,参与植物抵抗病虫害的过程中。通过田间调查和昆虫触角电生理分析,对这3种挥发物在稻田节肢动物群落和嗅觉行为水平上的作用进行了研究。结果发现水杨酸甲酯对害虫的虫口密度影响较明显,晚稻田中,水杨酸甲酯组的害虫数量(14.42头/板)显著高于其余各处理组。赤眼蜂的嗅觉电位随芳樟醇和茉莉酸甲酯浓度的升高而显著增大,浓度为1/50时赤眼蜂嗅觉电位的变化值显著高于浓度为1/5000时嗅觉电位变化值。     

水稻生产氮肥需要量与利用率试验研究

通过中稻氮肥效应试验,结果表明:在所设处理中,随施氮量的增加,水稻产量增加,并获得了氮肥效应函数方程Y=-0.65 X2+24.88 X+443.1.通过对植株吸氮量和氮肥利用率的比较,表明当地施氮水平(每667m2施纯氮10kg)的氮肥利用率较高,净产值和产投比较大,能增加稻谷产量和产值,适合当地当前中稻生产实际.     

Promoter of soybean early nodulin gene<Emphasis Type="Italic">enod2B</Emphasis> is induced by rhizobial Nod factors in transgenic rice

Nod factors, which are signaling molecules produced by Rhizobia, are the principal determinants of host specificity in Rhizobium-legume symbiosis. Nod factors can elicit a number of characteristic developmental responses in the roots of legumes, such as depolarization of the membrane potential in epidermal cells, specific expression of early nodulin genes and changes in the flux of calcium in root hairs, deformation of root hairs, cell division in the root cortex and formation of the nodule primordinm. Whether the rice plant can respond to signaling molecules (i.e. Nod factors) is an important question, as it could establish the potential for symbiotic nitrogen fixation in rice. The promoter of the soybean (Glycine max) early nodulin gene Gmenod2B fused to the β-glucuronidase (GUS) reporter gene was used as a molecular marker to explore whether Nod factors can be recognized by rice cells as signaling molecules. Transgenic rice plants harboring the chimeric gene Gmenod2BP-GUS were obtained via an Agrobacterium tumefaciens-mediated system. NodNGR factors produced by a broad-host-range Rhizobium strain NGR234(pA28) were used as probes to investigate the activity of the Gmenod2B promoter in rice. Our results showed that the early nodulin gene Gmenod2B promoter was induced by NodNGR factors in transgenic rice, and that it was specifically expressed in rice plant roots. Moreover, GUS gene expression driven by the Gmenod2B promoter in transgenic rice was regulated by nitrogen status. These findings indicated that rice possessed the ability to respond to Nod factor signals, and that this signal transduction system resulted in activation of the Gmenod2B promoter. Thus, we predict that the Nod-factor inducible nodulin expression system, which is similar to Rhizobium-legume symbiosis, may also exist in rice.     

不同施肥条件下巴西陆稻IAPAR9的养分吸收特性研究

巴西陆稻植株在移栽期——分蘖盛期的氮、磷、钾吸收量较少，分别占其吸收总量的17．1％，12．8％，13．0％，在分蘖盛期——孕穗期氮磷钾吸收量比重最大，分别占其吸收总量的50．1％，53．7％，48．7％左右。氮、磷、钾用量与土壤碱解氮、有效磷、速效钾，以及植株氮、磷、钾吸收总量呈极显著正相关，氮肥用量与植株钾素吸收总量亦呈极显著正相关。在中氮、磷、钾水平下，吸肥优势明显，物质生产效率较高。     

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。     

优质稻株的农艺性状与稻米品质关系的研究

测定桂花香等4个优质稻品种的农艺性状和稻米品质,进行相关分析和通径分析。农艺性状中的穗枝梗数和穗总粒数、穗实粒数之间呈极显著正相关,成穗率与穗长之间,穗复枝数与穗总数数之间呈显著正相关,而有效穗数与茎宽之间,千粒重与穗复枝数、穗实粒数均呈显著负相关。稻米品质中的糙米率与精米率呈极显著正相关,精米率与整精米率、蛋白质之间均呈显著正相关。整精米率、碱消值与直链淀粉含量之间,垩白率与糙米率、蛋白质之间均呈显著负相关,精米率与垩白率之间呈极显著负相关。农艺性状与稻米品质之间不呈显著相关。单株生产力对蛋白质含量的直接影响是负值,而生产力对直链淀粉含量的影响是正值,直接影响比间接影响大。     

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.     

Transformation of japonica rice with RHL gene and salt tolerance of the transgenic rice plant

Overexpression of the yeast HAL2 gene increases salt tolerance of yeast and plant. Rice HAL2-like (RHL) gene was introduced into a japonica rice cultivar HJ19 with Agrobacterium tumefaciens-mediated transformation. Transgenic plants in R0 generation were selected on the principle of GUS-positive, RHL gene PCR-positive and normal growth. Hygromycin-resistant plants of some transgenic lines in R1 generation increased salt tolerance during the seedling and booting stage, being less damaged in the cytomembrane and stronger in leaf tissue viability under salt stress during booting period. Southern analysis of transgenic lines tolerant to salt in R1 generation showed that the RHL gene expression cassette had been successfully integrated into rice genome. Moreover, gene engineering breeding methodology and really salt-tolerant rice cultivar were discussed.     

Molecular cloning of rice cDNAs related to pollen development using the subtraction hybridization technique

The meiotic stage of pollen mother cell is a very important stage in controlling the development and formation of pollen. In order to clone the rice cDNA(s) of this stage, a normal rice, Annong N and its thermosensitive mutant, Annong S-1 were used as the plant material. The mRNA has been extracted from the young panicle at the meiotic stage. By using the cDNA subtraction hybridization technique, three cDNA fragments, RP-1, RP-2 and RP-3 have been successfully cloned from Annong N. Northern blot analysis reveals that the mRNA of these three clones are expressed only in anthers, and not leaves. The mRNA levels of these clones are lower in anthers of Annong S-1 than in Annong N. Furthermore, the amount of mRNA extracted from anthers of Annong S-1 growing under high temperature (28℃) is lower than plants growing at lower temperature (25℃). Sequence analysis and homology search indicate that these three clones display no similarity to the current database. It is concluded that the three novel cDNA cloned are related to pollen development in rice.