High Temperature Modifies Resistance Performances of Rice Varieties to Brown Planthopper,Nilaparvata lugens(Stl)

To investigate the effect of temperature on the resistance characteristics of rice varieties with different resistance genes to brown planthopper(BPH),Nilaparvata lugens(Stl),the resistances of IR26(Bph1)and IR36(bph2)to BPH population in Hangzhou,China were monitored in greenhouse during September in 2007 and 2008 by using the standard seedling screening techniques(SSST)developed by the International Rice Research Institute(IRRI).Furthermore,the changes in resistance of IR26 and IR36 to BPH,soluble sugar and oxalic acid contents in 25-day-old rice plants of susceptible variety TN1 and resistant varieties IR26 and IR36 were detected at five temperatures(22℃,25℃,28℃,31℃ and 34℃).IR26 completely lost resistance both in greenhouse and at the five tested temperatures.IR36 still had moderate resistance at natural temperature,but its resistance decreased gradually when the temperature increased from 25℃ to 34℃,and fully lost its resistance at 31℃ and 34℃.The highest durable resistance of IR26 and IR36 were recorded at 25℃.The soluble sugar content in plants of the three tested rice varieties increased with temperature increase,and the oxalic acid content increased with the temperature increase at first,maximized at 25℃,and then declined.Two-way ANOVA indicated significant effects of temperature and rice variety on contents of soluble sugar and oxalic acid in rice plants.     

Bacterial Community in Different Populations of Rice Brown Planthopper Nilaparvata lugens （St~l）

The structures of bacterial communities in the brown planthopper （BPH） Nilaparvata lugens （St~l） from different geographic and resistant virulent populations were analyzed by using denatured gradient gel electrophoresis （DGGE）. Results showed that the bacterial communities in BPH nymph from the first to the fifth instars varied with nymphal growth and development. The bacterial communities in the first-instar BPH nymph were similar to those in adults. Nine geographic BPH populations were divided into three groups based on the cluster analysis of DGGE fingerprint. The first group was from the Philippines; the second group was from Thailand and Hainan, Yunnan and Zhejiang provinces of China; and the third group was from Vietnam and Guangxi, Hunan and Jiangxi provinces of China. BPH populations adapted to different resistant rice varieties. The BPH populations from Mudgo （with resistant gene Bphl） and ASD7 （with resistant gene bph2） differed with those of the susceptible rice variety TNI.     

Changes in Methamidophos Resistance and Fitness of Hybrids in Different Strains of Nilaparvata lugens

A resistant strain selected successively in the laboratory for 17 generations had 198.63-fold resistance to methamidophos. The resistant levels and fitness of progenies from the resistant strain and susceptible strain or field population were closer to those of the resistant strain than those of the susceptible strain or field population. The changes in the resistant levels of the hybrid were propitious to the resistance development, however, the changes of the fitness went to the contrary. The effects of the migration on the development of methamidophos resistance in Nilaparvata lugens were discussed in the aspects of the migration of Nilaparvata lugens, the resistant levels of progenies and the changes of the fitness.     

Virulence of the Populations of the Whitebacked Planthopper, Sogatella furcifera Reared on Different Resistant Rice Varieties

ARC and ND colony were obtained by rearing the whitebacked planthopper (WBPH), Sogatella furcifera(Horvath), exclusively on ARC10239 (ARC, carrying resistance gene Wbph2) and N‘Diang Marie (ND, carrying resistance gene WbphS) till 15 generations. Taichung Native 1 (TN1) and Rathu Heenati (RHT) served as susceptible and resistant check, respectively. The results of electronic recording revealed that duration of salivation and X-waveform of the two colonies on their corresponding hosts was short while the duration of phloem ingestion was long. The amount of honeydew excreted by ARC and ND colony did not differ on their corresponding host varieties from TN1 and was much higher than that of RHT. The number of eggs laid on their host varieties and TN1 were significantly higher than that on RHT. No distinct change was observed for these two colonies in term of percentage of developed eggs. The nymph survival rate of ND colony on its selection host was 45.0%, significantly different from that on TN1(71.4%0) and RHT(21.0%), while that of ARCcolony was 68. 3%, not significantly different from that on TN1(77.5%), but much higher than that on RHT(22.6%).The nymphal development duration of these two colonies on the corresponding hosts was not different from that on TN1, but significantly shorter than that on RHT. In brief, these two colonies had almost adapted to their corresponding host varieties based on feeding and oviposition, but the nymphal survival rate of ND colony was still low.     

Preliminary Study on Resistance of the Rice Stem Borer （Chilo Suppressalis） to Fipronil

By means of topical application, fipronil resistance was surveyed in the rice stem borer, Chilo suppressalis, from 12 diferent areas in east China from 2001 to 2004. The rice stem borers in most regions of Jiangsu and Anhui were still susceptible to fipronil. But in Wuxi, Jiangsu Province and Cixi, Zhejiang Province, their sensitivity became decreased （resistance ratios were 3.1 and 3.6, respectively）, and the medium level of resistance （resistance ratio was 21.2） was found in Cangnan, Zhejiang Province. So, it was still at the early stage for fipronil resistance development in this pest. Synergism experiments showed that piperonyl butoxide（PBO） had a little effect on both susceptible and resistant borers （synergism ratios were 1.1-1.2）. Though triphenyl phosphate （TPP） and diethyl meleate （DEM） had no effect on the susceptible borers, they had significant synergism on fipronil in the resistant population to fipronil （synergism ratios were 1.8 and 1.6, respectively）, indicating esterase and glutathion S-transferase may be involved in the resistance mechanism. Bioassay with currently used insecticides indicated that triazophos （because of high resistance）, trichlorphon and acephate had very low toxicity to resistant borers. But diazinon, pyridaphenthion, decamethrin and avermeatin showed high toxicity and had no cross resistance to fipronil, which could be considered as substitute insecticides in the resistance managment.     

Resistance Evaluation to Sheath Blight in Transgenic Rice Lines

Resistance of forty-one homozygous rice lines transformed with chitinase gene (RC24) and β-1,3 -glucanse gene β-1,3-Glu) to sheath blight was analyzed by inoculation. Among different lines, the resistance had significant differences according to the result by cluster analysis. The lines could be categorized into resistant, moderately resistant, moderately susceptible and susceptible types, while 92.1% of which belonging to moderately resistant or moderately susceptible type. For different resistant or susceptible lines, the resistance to rice sheath blight was remarkable correlated with the chitinase activity of transgenic lines except resistant type lines, in which enzyme activity coded by target gene was lower than moderately resistant type. The chitinase activity of transgenic lines tested at different time after inoculation or different organs of the same plant was uniform, which suggested that the expression of chitinase gene was constitutive in nature. Check varieties chitinase activity would change at different time after inoculation and reach a peak at sometime, but it had no difference at various pans of the same plant.     

Resistance Evaluation to Sheath Blight in Transgenic Rice Lines

Resistance of forty-one homozygous rice lines transformed with chitinase gene (RC24) and β-1,3 -glucanse gene (β-1,3-Glu) to sheath blight was analyzed by inoculation. Among different lines, the resistance had significant differences according to the result by cluster analysis. The lines could be categorized into resistant, moderately resistant, moderately susceptible and susceptible types, while 92.1% of which belonging to moderately resistant or moderately susceptible type. For different resistant or susceptible lines, the resistance to rice sheath blight was remarkable correlated with the chitinase activity of transgenic lines except resistant type lines, in which enzyme activity coded by target gene was lower than moderately resistant type. The chitinase activity of transgenic lines tested at different time after inoculation or different organs of the same plant was uniform, which suggested that the expression of chitinase gene was constitutive in nature. Check varieties‘ chitinase activity would change at different time after inoculation and reach a peak at sometime, but it had no difference at various parts of the same plant.     

Osa-mi R439 Negatively Regulates Rice Immunity Against Magnaporthe oryzae

Osa-miR439 is a rice-specific microRNA family. Here we showed that Osa-miR439 acted as anegative regulator in rice immunity against blast fungus Magnaporthe oryzae. Osa-miR439 differentiallyresponded to M. oryzae between susceptible and resistant rice accessions. The accumulation ofOsa-miR439 was constitutively more in the susceptible accession than in the resistant one. Transgeniclines overexpressing Osa-miR439a (OX439a) showed higher susceptibility associating with lower inductionof defense-related genes and less hydrogen peroxide (H2O2) accumulation at the infection sites than thecontrol plants. In contrast, transgenic lines expressing a target mimic of Osa-miR439 (MIM439) displayedcompromised susceptibility associating with increased H2O2 accumulation. Furthermore, we found thatthe expression of three predicted target genes was decreased in OX439a but increased in MIM439 incomparison to control plants, and this expression was differential in susceptible and resistant accessionsupon M. oryzae infection, indicating that Osa-miR439a may regulate rice blast resistance via these genes.Our results unveiled the role of Osa-miR439a in rice blast resistance and provided the potentiality toimprove the blast resistance via miRNA.     

Analyses of gene effects and inheritance of resistance to bacterial blight in rice

We analyzed the resistant inheritance of a japonica variety, Jia23, to two bacterial blight pathogen strains, KS—6—6 and Zhe 173, representing respectively the two predominant pathogenic types (Ⅱ and Ⅳ) in the rice cropping area along the Yangtze River Valley. Jla23 was crossed with susceptible varieties, Ewan8 and 7416. F_1 plants were backcrossed with both resistant and susceptible parents to produce B_1 (F_1×P_1) and B_2 (F_2×P_2). All generations were planted during the late—season on an isolated field from Jun to Oct in 1994. Thirty—day—old seedlings were transplanted individually with a spacing of 13cm between plants and 17cm between rows. Concentration of inoculum was adjusted to 10~9 cells/ml. Three fully expanded leaves per plant were inoculated with each type of strains by clipping     

Exploitation of genetic variation caused by space treatment in rice

Seeds of early indica rice varieties 6188, D6049, of late indiea rice varieties Minghui 63, Xiangge, Waiqi, 70321, and late japonica rice variety Beik 15 were carried by the retrieved satellite "921006" in order to investigate the comprehensive effects of the space factors. Treated seeds and untreated seeds (CK) were grown in the field. The results showed that although compared with the CK (which was very uniform) small differences on population uniformity, plant height, growth duration and seed setting rate were observed, no segregation was found in SP_1. Strong segregation occurred in SP_2 and SP_3 generations. Significant difference on the grain yield of Single plant was observed between SP_2 and CK, while the yield between     

Virulence Types of Magnaporthe oryzae to Hybrid Rice in Sichuan,China

A total of 638 isolates of rice blast(Magnaporthe oryzae) were isolated in 2002-2009 from different rice varieties in different regions of Sichuan,China and inoculated onto seven rice varieties(Lijiangxintuanheigu,IR24,Minghui 63,Duohui 1,Chenghui 448,Neihui 99-14 and RHR-1) to differentiate the virulence types of the fungus and trace the changes.The virulence to the seven varieties was respectively scored at 1,2,4,8,16,32 and 64.The total scores of individual M.grisea isolates which were the sum of scores infecting differential varieties could,in turn,be used for the nomenclature of the virulence types due to their accordance to the special virulence patterns.The 638 tested isolates were then differentiated into 56 different virulence types.Type 15 virulent to Lijiangxintuanheigu,IR24 and Minghui 63,and Type 127 virulent to all of the seven varieties were the most dominant virulence types respectively with the occurrence frequencies of 15.99% and 15.83%.Type 19 and other seven virulence types were not monitored during 2002-2009.Type 15 was the predominant virulence type in 2002,2003,2004 and 2007,whereas Type 127 had been the most dominant virulence type after 2005 except for the year 2007 when the province underwent severe drought.Five hundred and seven out of the 638 tested isolates were virulent to Minghui 63,and 89.58% of the 384 isolates virulent to either Duohui 1,Chenghui 448 or Neihui 99-14 were virulent to Minghui 63,which indicated the impact of the extensive plantation of hybrid rice Minghui 63 as the restorer line on the virulence evolution of M.oryzae in Sichuan.The virulence pattern of the dominant virulence types suggested that the acquiring of virulence to all the major resistant restorer lines was the main routes of the evolution in virulence of M.oryzae to hybrid rice in Sichuan.The virulence frequencies of the 638 tested isolates to IR24,Minghui 63,Duohui 1,Chenghui 448,Neihui 99-14 and RHR-1 were respectively 74.6%,79.5%,73.8%,37.0%,39.0% and 40.4%.The analysis for the sources of the different virulence type isolates indicated the pathogen on the newly released resistant varieties were stronger than conventional rice varieties which had become susceptible in the field since 1980s.     

Identification of a QTL for cold tolerance at early seedling stage via RFLP markers in rice （O. Sativa L. ）

Recently,beneficial genes in japonica and indica varieties were widely used from each other to promote the yield and insect and disease resistance,etc.in rice.Most indica varieties are susceptible to low temperature,and cold injury will affect the rice yield at various growth stages.     

Different Hypotheses for Resistance Loss of Rice Varieties to Magnaporthe oryzae

It is widely believed that a rice variety loses its resistance to rice blast after three to five years in commercial production due to the emergence of new Magnaporthe oryzae pathotypes(or physiological races)(Ou,1985).However,we believe that a resistant rice variety loses its resistance to M.oryzae not due to the emergence of new physiological races.We propose that pathogenic physiological races already exist in the natural environment,serving as potential physiological races.It is only because of the lack of appropriate environmental conditions,particularly the lack of a suitable rice host,on which such a race is unable to propagate widely and exists as an inferior race.However,when there is an appropriate rice host,the potential pathogenic physiological race can quickly proliferate to become the dominant physiological race,thus leading to the loss of blast resistance in resistant rice varieties.     

Genetic analysis of blast resistance On rice cultivars with wide—spectrum resistance

In recent years,indirect selection based ontightly linked genetic markers has been sug-gested to pyramid the resistance genes.How-ever,resistance parents used in the linkagestudies were usually near isogenic lines possessing a single resistance gene derived fromthe original resistance donor.Little was knownon the interaction among the genes at differentloci.In the present study,the genetic controlof blast resistance in two wide—spectrum re-vistance rice cultivars was analyzed.A susceptible indica varieties Xianfenglwas used as the female parent,and the blastresistance cultivars Tetep and Hongjiaozhanwere used as the male parents.The threeparental lines were inoculated with each of 20races of Pyricularia oryzae Cav at tilleringstage.The race ZC_(13) was selected to inoculatethe 161 F_3 lines of Xianfeng1/Tetep and 175 F,lines of Xianfengl/Hongjiaozhan.Eighteen in-dividuals of each line were inoculated.In both populations,all the individualsdemonstrated extreme reaction to the blast dis-ease.In the F_3 po     

Effect of Phytosulfokine-a on Agrobacterium-Mediated Transformation in Rice

Phytosulfokine- α （PSK- α ）, a biologically active peptide acting as a growth factor, plays a key role in cellular differentiation and proliferation. To test if PSK- α has some influence on agrobacterium-mediated transformation in rice, PSK-α at a series of concentrations was added into co-culture medium respectively. The results showed that PSK- α indeed affected the recovery of resistant calli and the transformation frequency of rice varieties Taipei 309 and Lijiangxintuanheigu, PSK- α at the concentration of 10 nmol/L could increase induction of resistant callus and efficiency of transformation, with a 11% and 4.9% top increase, respectively than the control. However, PSK- αat 200 nmol/L could inhibit the induction of the resistant calli. Further more, the effect of PSK-α on agrobacterium-mediated transformation is related with the concentration of 2, 4-D in selection medium. Higher induction rate of resistant calli was obtained from tissues treated with PSK- α plus 2 mg/L 2, 4-D.     

Tolerance evaluation of rice varieties to the brown plan-thopper,Nilaparvata lugens feeding

The brown planthopper, Nilaparvata lugens (Stal) (BPH) is one of the most important insect pests of rice in China and other east-southern Asian countries. Utilization of rice resistance varieties is one of the most e-conomic and effective ways for controlling N. lugens . The resistance of rice varieties declined due to the change of BPH biotype. The current resistance screening methods based on biotic reaction also may eliminate numerous rice varieties with high-yielding and/ or good grain quality which were not resistant to BPH but had strong tolerance to BPH.     

Effect of Temperature on Pollen Fertility in Inter-Subspecific Rice Hybrids

Seventeen rice varieties and hybrids of different types (indica, japonica, javanica, indica hybrid, japonica hybrid and inter-subspecific hybrid) were evaluated to determine the effect of temperature on pollen fertility in inter-subspecfic hybrids. The pollen fertility of inter-subspecific hybrids was greatly reduced when average daily temperature dropped to 22.0 - 23.0℃ at meiosis stage, and the extent of pollen fertility reduction varied greatly with respect to different hybrids. However, the pollen fertility reduction of indica and japonica hybrids and conventional varieties was not obvious under the same regime of temperature conditions. When the average daily temperature dropped to 20℃, the pollen development of conventional varieties and hybrids was also affected. Correlation analysis revealed that there existed a positive correlation between pollen fertility and average daily temperature. A significant difference (P＜0.01) was also found between the two correlation coefficients i.e. inter-subspecific hybrids and conventional varieties. Temperature at meiosis stage of pollen mother cell was a key factor in pollen developing, and the pollen fertility of inter-subspecific hybrids was more sensitive to low temperature than that of traditional variety.     

Effect of Temperature on Pollen Fertility in Inter-Subspecific Rice Hybrids

Seventeen rice varieties and hybrids of different types (indica, japonica, javanica, indica hybrid, japonica hybrid and inter-subspecific hybrid) were evaluated to determine the effect of temperature on pollen fertility in inter-subspecfic hybrids. The pollen fertility of inter-subspecific hybrids was greatly reduced when average daily temperature dropped to 22.0 - 23.0℃at meiosis stage, and the extent of pollen fertility reduction varied greatly with respect to different hybrids. However, the pollen fertility reduction of indica and japonica hybrids and conventional varieties was not obvious under the same regime of temperature conditions. When the average daily temperature dropped to 20℃, the pollen development of conventional varieties and hybrids was also affected. Correlation analysis revealed that there existed a positive correlation between pollen fertility and average daily temperature. A significant difference (P<0.01) was also found between the two correlation coefficients i.e. inter-subspecific hybrids and conventional varieties. Temperature at meiosis stage of pollen mother cell was a key factor in pollen developing, and the pollen fertility of inter-subspecific hybrids was more sensitive to low temperature than that of traditional variety.     

Physiological Macro-lesions Enhanced Resistance to Blast （Magnaporthe grisea） in Rice Near-isogenic Lines

Roll-leaf-1 （rl-1） and spot-leaf-1 （spl-1） were two near-isogenic lines, which were obtained after 3 to 4 backcrosses with early season indica rice Zhefu 802 as recurrent parent. Henna macro-lesions, referred as physiological or morphological markers, began to appear on leaves at 4.5- to 6.0-leaf stage. The rice seedlings were inoculated at 3.5-, 5.0- and 7.0-leaf stages with high pathogenic races Zhong A1 and Zhong B1 of Magnaporthe grisea, respectively. The resistance of rl-1, spl-1 and Zhefu 802 against blast was significantly different. The seedlings of Zhefu 802 at 3.5- to 7.0-leaf stage were susceptible to races Zhong A1 and Zhong B1 of M. grisea, whereas those of rl-1 and spl-1 at 3.5-, 5.0- and 7.0-leaf stages were susceptible, moderately resistant and resistant, respectively. These results suggested that the enhanced resistance of rl-1 and spl-1 related to the appearance of their morphological marker lesions. The experiment provided a basis for studying lesion mimic and hypersensitive response in association with disease resistance.