Analysis on Quantitative Trait Loci for Resistance to Rice False Smut under Different Environmental Conditions

A recombinant inbred line （RILs） population with 157 lines derived from an inter-subspecies cross of Daguandao （japonica）/IR28 （indica） by the single seed de-scent method was used to detect quantitative trait loci （QTLs） conferring resistance to rice false smut caused by Ustilaginoidea virens（Cooke） Takahashi in Nanjing and Yangzhou. The disease rate index of the two parents and 157 RILs caused by rice false smut were scored and the QTLs for rice false smut resistance were detected accordingly by QTL Cartographer software. Eight QTLs control ing false smut resis-tance were detected on chromosomes 1, 2, 4, 8, 10, 11 and 12, respectively, with the phenotypic variance of 8.6%-22.5%. There were five QTLs detected in Nanjing and Yangzhou, respectively, and only two QTLs were found in both two years, the phenotypic variation was explained by individual QTL ranged from 18.0% to 18.9% for these two QTLs, and the additive effects of these two QTLs contributed to the 8.0%-14.6% decrease of disease index and therefore the disease resistance increased. The direction of the additive effects at six loci qFsr1, qFsr2, qFsr8, qFsr10a, qFsr11 and qFsr12 coincided with that predicted by phenotypes of the parents, and the IR28 al eles at these loci had positive effect against rice false smut while the negative effects were found in Daguandao al eles at qFsr4 and qFsr10b. Both qFsr10a and qFsr11 should be useful in rice breeding for resistance to rice false smut in marker-assisted selection （MAS） program.

Resistance of Major and Backup Rice Cultivars against Ustilaginoidea virens in Anhui Province

[Objective] The aim was to study resistance of major and backup rice cul- tivars against Ustilaginoidea virens. [Method] Experiments and surveys were made on resistance of seventeen backup rice cultivars and some major cultivars in Anhui Province to identify resistance of different rice cultivars. [Result] Yanjing No.9, 80You 226, Tianxie No.l, A01 Xian, Lvjingnuo No.6 were moderate resistant; A03 Xian and Yangjing 636 were susceptible and the rest ten showed moderate susceptibility. Based on surveys on major rice cultivars, most of Liangyou rice series are suscept- able and novel Liangyou Xiang 4 enjoys resistance to certain extent. [Conclusion] The research provided references for resistance of rice against diseases in Anhui Province.

Effects of β-Amino Butyric Acid Induced Rice Blast Resistance on Reactive Oxygen Metabolism

[Objective] This study was to understand the effects of β-Amino butyric acid(abbreviated as BABA) induced rice blast resistance on reactive oxygen metabolism. [Method] Using the cultivar Chaochan 2 that is highly susceptible to disease as experimental material, the changes of catalase(CAT), and superoxide dismutase(SOD) and MDA activities in rice treated by BABA were investigated. [Result] In rice plants treated by BABA, the activities of CAT and SOD increased, meanwhile the MDA content also rose to some extent, resulting in the disease resistance to rice blast. [Conclusion] By influencing reactive oxygen metabolism, BABA endows rice plants with resistance to rice blast. BABA is safe to environment and has highly resistance-inducing capacity, it could be generalized in production.

Genotypic Analysis of Rice Blast Resistance Genes Pi-ta and Pi-b in Japonica Rice Varieties and Lines in Jiangsu Province

Pi-ta and Pi-b, the first cloned rice blast resistant genes, have been wide- ly used in rice blast resistance breeding for their lasting and stable resistance. To define the distribution of Pi-ta and Pi-b in japonica rice in Jiangsu, the genotypes of resistance genes Pi-ta and Pi-b in 40 varieties and 665 new lines were detected using functional markers of Pi-ta/pi-ta and Pi-b^pi-b alleles. The results showed that the resistance alleles of Pi-ta and Pi-b were widely spread in japonica rice varieties, and the distribution frequency of Pi-b was higher than that of Pi-ta. Most of the Lianjing serial varieties didn＇t carry the two resistance genes, but the two resistance genes were widely distributed in Wujing serial varieties. There was no significant dif- ference in distribution frequency of Pi-ta between new lines and commercial vari- eties. However, the distribution frequency of Pi-b in new lines was higher than that in commercial varieties. It was indicated that artificial selection was conducive to the improvement of distribution frequency of Pi-b in rice varieties. Among the 4 genotypes, the distribution frequency of pi-taJPi-b was highest （60.0%）, followed by Pi-ta/ Pi-b （33.5%） and pi-ta/pi-b （3.9%）. The frequency of Pi-taJpi-b was lowest, account- ing for only 2.6%. In terms of source of resistance genes in the four combinations, the resistant allele Pi-ta might be from parents of Wuxiangjing14, Wujing15 or Nanjing44, and Pi-b might come from parents of Wujing13, Wuxiangjing14, Wujing15 or Nanjing44. The analysis on the genotypic frequencies in offspring of the rice vari- eties showed that the resistance genotype of Pi-ta/Pi-b had the highest frequency in the cross combination of Nanjing44//Wujing13/Kantou194.

Improvement of Resistance to Rice Blast in Zhenshan 97 by Molecular Marker-aided Selection

Fungi blast is one of the most serious diseases of rice worldwide. Breeding resistant varieties have been proved to be the most effective and economical means to control the disease. This paper describes the molecular marker-assisted selection (MAS) procedure for a broad-spectrum blast resistant gene Pi1 integrated into an elite hybrid maintainer line, Zhenshan 97. A simple sequence repeat (SSR) based on molecular marker-aided selection system for Pi1 segment was established. Using a backcross population and a blast isolate F1829, Pi1 gene was mapped on the top of chromosome 11 between markers RZ536 and RM144, with a distance of 9.7 cM and 6.8 cM, respectively. Seventeen families derived from the recurrent parent Zhenshan 97 were obtained with homozygous Pi1 gene. The background of the 17 families was identified with inter simple sequence repeat (ISSR) amplification, the highest recovery of the Zhenshan 97 genetic background was 97.01% after the assay of 167 polymorphic bands.

Effect of Morphological Traits on Sheath Blight Resistance in Rice

Sheath blight, caused by the Rhizoctonia solani Kuhn, is one of the most serious diseases of rice (Oryza sativa L.). The relationship between morphological traits and sheath blight resistance was investigated in 1999 and 2000 by using a segregant population consisting of 240 inbred recombination lines, derived from an elite combination of Zhenshan 97 X Minghui 63. Partial correlation analysis between phenotypic data (morphological characteristics and heading date (HD)) and sheath blight ratings (SBR) showed that only one trait, plant compactness, was significantly correlated with resistance to sheath blight in both 1999 and 2000, eight traits, such as plant height, heading date, and penultimate leaf angle, were significantly correlated with SBR in either of the two years, and the remaining 10 morphological traits were not consistently associated with SBR in the two years. A major quantitative trait locus (QTL) controlling plant compactness was found to be in the site on chromosome 9 adjacent to the region responsible for qSB9, a major QTLs conferring sheath blight resistance. One out of the three QTLs contributing to a penultimate leaf angle was mapped approximately on the same region as another sheath blight resistance QTL, qSB5, located on chromosome 5. Whereas, no QTLs underlying most other traits was detected on the chromosomal region correlated with sheath blight resistance QTL. The results in the present study suggested that the morphological traits were not the main factors responsible for the SBR separation among the recombinant inbred lines (RILs) population. Nevertheless, some morphological characteristics had some indirect influence on expression of sheath blight resistance genes through altering a microclimate on paddy field so as to influence the infection of the pathogen and development of the disease. An efficient approach in resistance breeding to sheath blight was recommended by pyramiding major QTLs for sheath blight resistance and selecting those morphological traits that favor resistance gene expression.

Screening of Rice Stripe Virus(RSV)-resistant Germplasm from Landraces of Rice(Oryza sativa L.)

[Objective] The aim of this study was to screen rice strip virus （RSV）-resistant landraces. [Method] The resistance of 119 rice landraces to rice stripe virus was identified in field spontaneously infected with smal plant-hopper. [Result] There were 55 landraces resistant to rice strip disease in 56 indica rice landraces, but on-ly two resistant to rice strip disease in 63 japonica rice landraces. [Conclusion] The results revealed that there were abundant rice landscapes resistant to RSV in Chi-na, and these varieties can be used to develop more genes resistant to RSV.

Mapping QTLs Associated with Sheath Blight Resistance Using Chromosome Segment Substitution Lines of Rice(Oryza sativa L.)

In this study, a population of 119 chromosome segment substitution lines （CSSLs） derived from backcross between indica 9311 and japonica Nipponbare was employed to map quantitative trait loci （QTL） associated with sheath blight resis-tance in rice with toothpick inoculation method. A total of three sheath blight resis-tance-associated QTLs （qsb8-1, qsb8-2 and qsb8-3） were identified, which were lo-cated on adjacent molecular markers RM3262, RM5485 and RM3496 of chromo-some 8; the genetic interval was 81.7cM-91.7cM, 91.7cM-108.1cM and 108.1cM-119.6cM, respectively. The additive effect of qsb8-2 was negative, indicating that sheath blight resistance of susceptible parent harboring qsb8-2 fragment was en-hanced; additive effects of qsb8-1 and qsb8-3 were positive, indicating that sheath blight resistance of susceptible parent harboring qsb8-1 and qsb8-3 fragments was reduced.

Mapping of QTLs for Sheath Blight Resistance Using Recombinant Inbred Lines of Rice(Oryza sativa L.)

A recombinant inbred line （RIL） population composed of 157 lines derived from an inter-subspecific hybrid of Daguandao/IR28 by the single seed descent method was used as materials, and the quantitative trait loci （QTLs） coffering the resistance to sheath blight in the 157 RILs and the parents were detected using the toothpick inoculation method. The disease indexes of rice sheath blight in the two parents and 157 RILs were scored and the QTLs responsible for rice sheath blight resistance were detected accordingly by QTL Cartographer software. The results showed that a total of 4 QTLs （qsbl, qsb2, qsb5-1, qsb5-2） conferring sheath blight resistance were detected on chromosomes 1, 2 and 5, and their variance explained ranged from 10.41% to 36.92%. The additive effect of qsb5-1 was negative, indicat- ing that the QTLs derived from donor parent IR 28 could enhance the resistance to sheath blight. However, the additive effects of qsbl, qsb2 and qsb5-2 were positive, indicating that the QTLs derived from donor parent Daguandao weakened the resis- tance to sheath blight.

Resistance Evaluation of Part of Leading Rice Varieties to Rice Blast in Guangxi Province

In order to determine the resistance of leading rice varieties to rice blast in Guangxi, three natural induction rice blast nurseries were established in Cenxi, Sanjiang and Yongfu. The resistance of the 45 leading rice varieties in Guangxi to rice blast was determined under conditions of spray inoculation and natural induction at seedling stage. The results showed that resistance frequencies of the 45 leading rice varieties ranged from 8.33% to 94.44%. Among the 45 tested varieties, there were 8 varieties of which the resistance frequencies were all higher than 80%, accounting for 17.78%; there were 11 varieties of which the resistance frequencies ranged from 50% to 80%, accounting for 24.44%; the resistance frequencies of the remaining varieties were all lower than 50%, accounting for 57.78% of the total tested varieties. The twoconsecutive-year determination results showed none of the 45 leading rice varieties showed resistance to rice blast. In 2010, there were 26, 12 and 7 varieties that were moderately sensitive, sensitive and greatly sensitive to rice blast respectively, accounting for 57.78%, 26.67% and 15.56% of the total tested varieties. In 2011, there were 16, 23 and 12 varieties that were moderately sensitive, sensitive and greatly sensitive to rice blast respectively, accounting for 35.56%, 51.11% and 13.33%. The overall resistance of the 45 leading rice varieties trended to be decreased year by year. So in rice production, they should be selected properly.

Analysis of Quantitative Trait Loci for Pathogenic Strain Pi-1 of Resistance to Rice False Smut in Rice(Oryza sativa)

In order to identify the resistant gene of rice false smut in rice, a recombi- nant inbred line （RILs） population with 157 lines derived from an inter-subspecies cross of Daguandao/IR28 by the single seed descent method was used to detect quantitative trait loci （QTLs） conferring resistance to strain Pi-1 of rice false smut caused by Usti/aginoiclea virens （Cooke） Takahashi in 2012 and 2013. The disease rate indexes of the two parents and 157 RILs caused by the strain Pi-1 of rice false smut were scored and the QTLs for rice false smut resistance were detected accordingly by QTL Cartographer software. Seven QTLs controlling false smut re- sistance were detected on chromosomes 2, 7, 8, 11 and 12, respectively, with the phenotypic variance of 8.5%-17.2%. There were four QTLs detected in 2012 and 2013, respectively, and only one QTL was found in both two years, the phenotypic variation explained by this individual QTL was 13.5% and 17.2%, and the additive effects of this QTL contributed to the 9.9% and 14.3% decrease of disease index and therefore the disease resistance increased. The direction of the additive effects at five loci qFsr2a, qFsr8a, qFsr8b, qFsr11 and qFsr12 coincided with that predicted by phenotypes of the parents, and the IR28 alleles at these loci had positive effect against rice false smut while the negative effects were found in Daguandao alleles at qFsr2b and qFsr7. The qFsr11 should be useful in rice breeding for resistance to rice false smut in marker-assisted selection （MAS） program.

Analysis on Resistance of Rice Cultivar Lianjing 7 to Rice Black-streaked Dwarf Disease

To analyze the resistance of Lianjing 7 against rice black-streaked dwarf disease and find the molecular markers closely linked with the resistance gene, an F2：3 population derived from an RBSDD-resistant cultivar Lianjing 7 and an RBSDD-susceptible cultivar Peiai 64, was evaluated using natural infection methods and artificial inoculation by scoring the disease severity rating. The results showed Lianjing 7 was resistant to RBSDD, with the incidence between 5.5% and 12.25% in all tests. For the parents of Lianjing 7, only Zhongjingchuan-2 was resistant to RBSDD. Besides this, Lianjing 7 assumed an antixenosis and antibiosis score similar with susceptible variety Wuyujing 3, which indicates that so RBSDD-resistance of Lianjing 7 is mainly caused by the resistance specifically to RBSDV. In the F2：3 population of Lianjing 7/Peiai 64, the disease severity rates showed a continuous distribution with transgressive segregation, it indicated the resistance of Lianjing 7 was controlled by multi-genes. The marker RM287 was determined closely linked with the resistance locus via BSA method. To conclude, Lianjing 7 has a high resistance to RBSDD, which is conferred with by Zhongjingchuan-2, and the marker RM287 should be useful for breeding novel RBSDD-resistant cultivars via marker-assisted selection （MAS）.

Constitutive expression of pathogen-inducible OsWRKY31 enhances disease resistance and affects root growth and auxin response in transgenic rice plants

WRKY transcription factors have many regulatory roles we isolated a rice WRKY gene （OsWRKY31） that is induced in response to biotic and abiotic stresses. In this study, by the rice blast fungus Magnaporthe grisea and auxin. This gene encodes a polypeptide of 211 amino-acid residues and belongs to a subgroup of the rice WRKY gene family that probably originated after the divergence of monocot and dicot plants. OsWRKY31 was found to be localized to the nucleus of onion epidermis cells to transiently express OsWRKY31-eGFP fusion protein. Analysis of OsWRKY31 and its mutants fused with a Gal4 DNA-binding domain indicated that OsWRKY31 has transactivation activity in yeast. Overexpression of the OsWRKY31 gene was found to enhance resistance against infection with M. grisea, and the transgenic lines exhibited reduced lateral root formation and elongation compared with wild-type and RNAi plants. The lines with overexpression showed constitutive expression of many defense-related genes, such as PBZI and OsSci2, as well as early auxin-response genes, such as OslAA4 and OsCrll genes. Furthermore, the plants with overexpression were less sensitive to exogenously supplied IBA, NAA and 2,4-1） at high concentrations, suggesting that overexpression of the OsWRKY31 gene might alter the auxin response or transport. These results also suggest that OsWRKY31 might be a common component in the signal transduction pathways of the auxin response and the defense response in rice.

Molecular Cloning of Early-expressed cDNAs from Rice in Response to Infection by Rice Blast Fungus Magnaporthe grisea

Compatible and incompatible reactions in rice plants ( Oryza sativa L. cv. Shenxianggen No.4) were resulted from inoculation with two different virulent races of rice blast fungus ( Magnaporthe grisea (Hebert) Barr), and thus an effective infecting system was established between rice plants and the rice blast pathogen. Two cDNA clones that showed induced and temporal patterns in expression in the very early stage in response to infection of the fungus were obtained from the plants by use of differential display. Of the two cDNA clones, Fastresp_a was induced to express in both compatible and incompatible interactions although it was expressed earlier in the former reaction. The second one, Fastresp_b , was only expressed in incompatible interaction. Southern blot analysis of the rice genomic DNA indicated that both of the two clones were from genome of the plant. No significant homology to the two genes was found from the rice gene database. This suggested that they were novel genes in rice and may play important roles in rice resistant response to infection of rice blast fungus.

Alpha-picolinic acid,a fungal toxin and mammal apoptosis-inducing agent,elicits hypersensitive-like response and enhances disease resistance in rice

Alpha-picolinic acid (PA),a metabolite of tryptophan and an inducer of apoptosis in the animal cell,has been reported to be a toxin produced by some of plant fungal pathogens and used in screening for disease resistant mutants. Here,we report that PA is an efficient apoptosis agent triggering cell death of hypersensitive-like response in planta. Confirmed by Fluorescence Activated Cell Sorter (FACS),rice suspension cells and leaves exhibited programmed cell death induced by PA. The PA-induced cell death was associated with the accumulation of reactive oxygen species that could be blocked by diphenylene iodonium chloride,indicating that the generation of reactive oxygen species was NADPHoxidase dependent. We also demonstrated the induction of rice defense-related genes and subsequent resistant enhancement by PA against the rice blast fungus Magnaporthe grisea. Hence,it was concluded that the PA-stimulated defense response likely involves the onset of the hypersensitive response in rice,which also provides a simple eliciting tool for studying apoptosis in the plant cell.

Cloning and Expression of the Serine Carboxypeptidase Gene in Zea mays and Its Antifungal Activity against Rhizoctonia solani

The authors cloned and identified a new maize serine carboxypeptidase gene named ZmSCP from R15 inbred lines seedlings which were induced by Rhizoctonia solani AGI-IA. ZmSCP encodes a 332 amino acid protein with a predicted molecular mass of 36.5 kDa and pI of 4.75. Phylogenetic analysis revealed that ZmSCP showed closer kinship with Oryza sativa and sorghum, which belong to the same evolutionary branch. Amino acid sequence analysis revealed that there are four types of amino acids in ZmSCP, the percentages of them are 43.1%, 26.9%, 13.9% and 13.1%. The authors subsequently purified the recombinant protein which expressed in Escherichia coli BL21 and analyzed its antimicrobial activities in vitro. Results showed that the recombinant protein inhibited hyphal growth of Rhizoctonia solani. The study suggests that the expression of ZmSCP is closely related to maize sheath blight resistance caused by Rhizoctonia solani. Further, the antifungal activity showed that ZmSCP may play at role in the disease resistance response.

Characterization of a disease susceptibility locus for exploring an efficient way to improve rice resistance against bacterial blight

Bacterial blight caused by Xanthomonas oryzae pv.oryzae(Xoo) is the most harmful bacterial disease of rice worldwide.Previously,we characterized major disease resistance(MR) gene xa25,which confers race-specific resistance to Xoo strain PXO339.The xa25 is a recessive allele of the SWEET13 locus,but SWEET13's interaction with PXO339 and how efficiently using this locus for rice breeding still need to be defined.Here we show that the SWEET 13 allele from rice Zhenshan 97 is a susceptibility gene to PXO339.Using this allele's promoter to regulate xa25 resulted in disease,suggesting that the promoter is a key determinant in SWEET13 caused disease in Zhanshan 97 after PXO339 infection.PXO339 transcriptionally induces SWEET13 to cause disease.Partial suppressing SWEET13 expression leads to a high level of resistance to PXO339.Thus,the transcriptionally suppressed SWEET13 functions as xa25 in resistance to PXO339.Hybrid rice is widely grown in many countries.However,recessive MR genes have not been efficiently used for disease resistance breeding in hybrid rice production for both parents of the hybrid have to carry the same recessive gene.However,the suppressed SWEET 13 functions dominantly,which will have advantage to improve the resistance of hybrid rice to xa25-incomptible Xoo.