Tolerance of longer-term partial stagnant flooding is independent of the SUB1 locus in rice

Longer-term partial stagnant flooding, particularly when it occurs following transient complete submergence causes severe damage to modern rice varieties. Progress was made in developing varieties tolerant of complete submergence through transfer of the Submergence-1 (SUB1) gene into popular varieties. However, SUB1 may not be effective under partial stagnant flooding (SF), as the new varieties may not elongate and continue growth when fully or partially submerged because of the SUB1-mediated suppression of elongation. We tested a set of rice genotypes, including a pair of near-isogenic lines (NILs), Swarna and Swarna-Sub1, under either SF or SF following complete submergence of 12 d. Swarna-Sub1 had higher survival and yield than Swarna following 12 d of submergence, but survival and grain yield of all lines decreased substantially when SF of 15-30 cm followed complete submergence, with the sensitive lines experiencing greater reductions in growth and yield. This suggests the importance of combining SUB1 with tolerance of SF in areas where both stresses are expected during the season. Swarna and Swarna-Sub1 are more sensitive to long-term partial SF than IR49830 and IR42 because of their short stature, and Swarna-Sub1 showed slightly higher reduction in tillering than did Swarna when subjected to deeper SF alone, possibly because of further inhibition of elongation by SUB1 if induced in submerged tissue. The results suggest that tolerance of these genotypes to SF depends less on SUB1 introgression and more so on the genetic background of the recipient genotype, with better performance of the genotypes that are inherently taller, such as IR42 and IR49830. The SUB1 donor landrace FR13A and its derivative breeding line IR49830 had better survival and relatively less reduction in grain yield under SF following complete submergence and under longer-term partial SF, indicating that these genotypes may have genes other than SUB1 for submergence and SF tolerance. For better adaptation to prolonged SF, SUB1 should be introgressed into genotypes that tolerate partial SF through better tillering ability and taller shoots, because SUB1 may not be effective in shorter genotypes, as it promotes survival of submerged plants by hindering shoot elongation to conserve energy reserves. Varieties combining tolerance of prolonged SF with SUB1 will have broader adaptation in flood-prone areas and greater impacts on yield stability.     

Changing allele frequencies associated with specific resistance genes to leaf blast in backcross introgression lines of Khao Dawk Mali 105 developed from a conventional selection program

Key genomic regions associated with blast resistance against a broad spectrum of isolates could be identified in backcross introgression lines developed by conventional breeding program. In this article, eighty three BC₃F₂ backcross introgression lines (BILs) derived from a cross between IR68835-98-2-B-2-1-1 (a broad spectrum blast resistance variety) and KDML105 (a susceptible variety) were developed by phenotypic selection against a mixture of six virulent blast isolates (MXL) that are widely spread out in the rainfed lowland of the North and Northeast of Thailand. The resistance spectrum of the BILs was assessed by inoculating with 12 different Magnaporthe oryzae isolates that showed differential responses on the parental cultivars and the MXL that was used for phenotypic selection. All BILs showed highly resistant reactions (low disease score) to the MXL and to two blast isolates THL48 and THL149. Four markers, i.e., RM246, RM241, RM303 and RM164 completely favoring the IR68835 allele (shifting of allele frequencies from KDML105 to IR68835) were identified on chromosomes 1, 4 and 5, respectively. Therefore, these markers could be linked to the resistance genes functioning against the MXL, THL48 and THL149. Furthermore, significant shifting of alleles was identified at six markers located on chromosomes 2, 4, 8, 9 and 12. Seven DNA markers linked to specific resistance genes were identified, in which allele from IR68835 at 6 markers, i.e., RM6, RM205, RM211, RM252, RM273 and RM342 reduced disease score (DS), against blast isolates THL16, THL329, THL458, THL831, THL868 and THL96036 while allele from KDML105 at RM208 (Pi-kd on chromosomes 2) reduce DS against THL84, THL191, THL557 and THL1108. In this study, the use of DNA markers enabled the identification of specific resistance genes in the backcross breeding materials developed from routine rice breeding program through the conventional phenotypic selection. In this experiment, the usefulness of breeding materials from conventional breeding program in identifying genes corresponding to the selection was illustrated. Linked markers and their genomic location provide necessary information for further use of marker-assisted selection to improve blast resistance in rice breeding program.     

Improving Rice Blast Resistance by Mining Broad-Spectrum Resistance Genes at Pik Locus

Magnaporthe oryzae is known for its genetic diversity and pathogenic variability, leading to rapid breakdown of resistance in rice. Incorporating multiple broad-spectrum blast resistance genes into rice cultivars would extend disease resistance longevity. Effective resistance breeding in rice therefore requires continual enrichment of the reservoir of resistance genes and alleles. We conducted a large-scale screen of rice blast resistance in about 2 000 rice accessions. Among them, 247 accessions showed at least medium resistance to the natural infection of rice blast and 7 novel Pik alleles were identified from them. Variations in gene sequences were then correlated with the phenotypic trait to enable the identification of favorable alleles. Among the seven novel Pik alleles, the resistant rate of Pik-R0/ME/7017 donors was greater than 80%, and the disease score was less than 3. Through molecular marker-assisted backcross breeding, we successfully transferred the three Pik alleles, Pik-R0/ME/7017, into an elite cultivated line Kongyu 131 to obtain BC₃F₂ lines, which showed enhanced resistance to rice blast compared with the recurrent parent. Assessment of these near-isogenic lines in the greenhouse using 31 isolates of M. oryzae from Heilongjiang Province of China revealed that the resistant levels of the BC₃F₂ lines with Pik-R0/ME/7017 were significantly higher than those of the established cloned resistance genes Pik-m and Pi1. Exploring such alleles will enrich our gene library for resistance to rice blast.     

水稻抗性基因Pi对福建省稻瘟病菌优势菌群的抗性分析

【目的】抗病品种的合理利用和布局是实现水稻抗瘟持久化的关键因子之一。为明确水稻24个抗性基因在福建省的抗病性及其应用前景,【方法】首先分别采用全国稻瘟病菌生理小种鉴别品种和CO39近等基因系来鉴定2012-2015年间从福建省各稻区种植的普感品种丽江新团黑谷上采集的347株稻瘟病菌单孢菌株的生理小种类型和致病型,再测定24个抗性基因对福建省稻瘟病菌的抗性。【结果】根据全国稻瘟病菌生理小种鉴别品种对菌株的抗感反应,可将供试稻瘟病菌的生理小种划分为6个群36个生理小种,其中ZA、ZB和ZC为主要种群,ZC15、ZD7和ZB15为优势生理小种。根据CO39近等基因系的接种结果,将供试稻瘟病菌划分为17个致病类型,其中I34.1为优势致病型。供试的24个抗性基因对347株福建省稻瘟病菌菌株的抗性频率不同,抗谱为9.80%~89.91%。其中,Pi-k~m、Pi-7(t)、Pi-9(t)、Pi-k~p、Pi-k、Pi-k~h、Pi-z~5和Pi-ta(1)等8个抗性基因的抗病性较强,抗谱均高于70.00%。【结论】说明这8个抗性基因在福建省具有较好的应用前景,育种时可以考虑联合利用这些抗性基因。同时,实验结果表明,这8个抗性基因对主要生理小种的抗谱和主要致病型的抗谱均值高于69.00%,与其对所有测试菌株的抗谱吻合。说明利用稻瘟病菌的优势种群或优势致病型来鉴定水稻品种的抗病性是可靠的。     

Identification of resistant germplasm containing novel resistance genes at or tightly linked to the <Emphasis Type="Italic">Pi2/9</Emphasis> locus conferring broad-spectrum resistance against rice blast

Background

The rice Pi2/9 locus harbors multiple resistance (R) genes each controlling broad-spectrum resistance against diverse isolates of Magnaporthe oryzae, a fungal pathogen causing devastating blast disease to rice. Identification of more resistance germplasm containing novel R genes at or tightly linked to the Pi2/9 locus would promote breeding of resistance rice cultivars.

Results

In this study, we aim to identify resistant germplasm containing novel R genes at or tightly linked to the Pi2/9 locus using a molecular marker, designated as Pi2/9-RH (Pi2/9 resistant haplotype), developed from the 5′ portion of the Pi2 sequence which was conserved only in the rice lines containing functional Pi2/9 alleles. DNA analysis using Pi2/9-RH identified 24 positive lines in 55 shortlisted landraces which showed resistance to 4 rice blast isolates. Analysis of partial sequences of the full-length cDNAs of Pi2/9 homologues resulted in the clustering of these 24 lines into 5 haplotypes each containing different Pi2/9 homologues which were designated as Pi2/9-A5, ?A15, ?A42, ?A53, and -A54. Interestingly, Pi2/9-A5 and Pi2/9-A54 are identical to Piz-t and Pi2, respectively. To validate the association of other three novel Pi2/9 homologues with the blast resistance, monogenic lines at BC₃F₃ generation were generated by marker assisted backcrossing (MABC). Resistance assessment of the derived monogenic lines in both the greenhouse and the field hotspot indicated that they all controlled broad-spectrum resistance against rice blast. Moreover, genetic analysis revealed that the blast resistance of these three monogenic lines was co-segregated with Pi2/9-RH, suggesting that the Pi2/9 locus or tightly linked loci could be responsible for the resistance.

Conclusion

The newly developed marker Pi2/9-RH could be used as a potentially diagnostic marker for the quick identification of resistant donors containing functional Pi2/9 alleles or unknown linked R genes. The three new monogenic lines containing the Pi2/9 introgression segment could be used as valuable materials for disease assessment and resistance donors in breeding program.

     

Improvement of Upland Rice Variety by Pyramiding Drought Tolerance QTL with Two Major Blast Resistance Genes for Sustainable Rice Production

Varalu is an early maturing rice variety widely grown in the rainfed ecosystem preferred for its grain type and cooking quality. However, the yield of Varalu is substantially low since it is being affected by reproductive drought stress along with the blast disease. The genetic improvement of Varalu was done by introgressing a major yield QTL, qDTY_12.1, along with two major blast resistance genes i.e. Pi54 and Pi1 through marker-assisted backcross breeding. Both traits were transferred till BC₂ generation and intercrossing was followed to pyramid the two traits. Stringent foreground selection was carried out using linked markers as well as peak markers (RM28099, RM28130, RM511 and RM28163) for the targeted QTL (qDTY_12.1), RM206 for Pi54 and RM224 for Pi1. Extensive background selection was done using genome-wide SSR markers. Six best lines (MSM-36, MSM-49, MSM-53, MSM-57, MSM-60 and MSM-63) having qDTY_12.1 and two blast resistance genes in homozygous condition with recurrent parent genome of 95.0%-96.5% having minimal linkage drag of about 0.1 to 0.7 Mb were identified. These lines showed yield advantage under drought stress as well as irrigated conditions. MSM-36 showed better performance in the national coordinated trials conducted across India, which indicated that improved lines of Varalu expected to replace Varalu and may have an important role in sustaining rice production. The present study demonstrated the successful marker-assisted pyramiding strategy for introgression of genes/QTLs conferring biotic stress resistance and yield under abiotic stress in rice.     

Development of STS markers and establishment of multiplex PCR for Glu-A3 alleles in common wheat (Triticum aestivum L.)

Low-molecular-weight glutenin subunits (LMW-GS) play a key role in determining the processing quality of the end-use products of common wheat. The objectives of this study were to identify genes at Glu-A3 locus, develop the STS markers, and establish multiplex PCR with the STS markers for Glu-A3 alleles. Gene-specific PCR primers were designed to amplify six near-isogenic lines (NILs) and Glenlea with different Glu-A3 alleles (a, b, c, d, e, f and g) defined by the protein electrophoretic mobility. Three Glu-A3 genes with complete coding sequence were cloned, designated as GluA3-1, GluA3-2 and GluA3-3, respectively. Seven dominant allele-specific STS (sequence tagged sites) markers were designed based on the SNPs (single nucleotide polymorphisms) among different allelic variants for the discrimination of the Glu-A3 protein alleles a, b, c, d, e, f and g. Four multiplex PCRs were established including Glu-A3b + Glu-A3f, Glu-A3d + Glu-A3f, Glu-A3d + Glu-A3g, and Glu-A3b + Glu-A3e. These markers and multiplex-PCR systems were validated on 141 CIMMYT wheat varieties and advanced lines with different Glu-A3 alleles, confirming that they can be efficiently used in marker-assisted breeding.     

Relationship between Shoot Elongation and Dry Matter Weight During Submergence in Oryza sativa L. and O. glaberrima Steud. Rice Cultivars

Abstract

Rice plants are damaged by flash floods with a rapid increase in water level caused by a heavy rain. However, rice plants cope with the flash floods either by an “escape strategy” involving rapid shoot elongation or by a “quiescence strategy” involving surviving under water with minimal activity. As we found in previous experiments, Saligbeli cultivar adapted well to flash floods through rapid shoot elongation. To understand the vigorous growth process during submergence, we studied the relationship between shoot elongation and changes in dry matter weight (DMW) during submergence. O. glaberrima Steud. cv. Saligbeli and O. sativa L. cv. Ballawé and IR 49830-7-1-2-2 were used. Saligbeli and Ballawé exhibit shoot elongation, and IR 49830-7-1-2-2 exhibits flash-flood tolerance due to the presence of the Sub-1 gene. Twelve-day-old seedlings were submerged for 7 days and the plant length and DMW were measured. The plant length ratio of submerged to control plants in Saligbeli was higher than that of other cultivars during 2-6 days of submergence but IR 49830-7-1-2-2 shoot elongation was inhibited by submergence. In all three cultivars, the elongation of the developing leaf sheath conferred shoot elongation during submergence. The plant length of all submerged plants showed a strong positive correlation with DMW of the leaves developed during submergence. Submerged Saligbeli and Ballawé showed strong negative correlations between DMW of the leaves developed before and during submergence (r = –0.786 for Saligbeli and –0.772 for Ballawé , P < 0.05), suggesting that the enhancement of shoot elongation during submergence is accomplished by using dry matter of the leaves developed before submergence. However, the correlation was not observed in the submerged IR 49830-7-1-2-2. Further details from studies using isotopes are also needed to understand the plant growth during submergence.     

江苏省稻瘟病菌群体分布及优势小种的毒力研究

用中国7个统一鉴别品种对2000~2002年江苏省5大稻区采集的342个稻瘟病菌单孢分离菌株进行了小种鉴定。结果表明3年累计出现7群30个小种,其中ZG1为优势小种,3年的频率分别为65.00%、56.90%和60.38%,ZB群和ZC群生理小种也是重要的小种类型,有较高的出现频率。用130个ZG1小种的菌株对日本13个已知抗性基因品种进行毒力测定,出现42种类型毒力,其中30.77％的ZG1小种的菌株同时对品种新2号（Pita-k[sup]s[/sup],Pi-sh）、K1（Pi-ta）、Pi-4号（Pi-ta[sup]2[/sup],Pi-sh）致病,为优势毒力类型, 表明品种新2号（Pita-ks,Pi-sh）、K1（Pi-ta）、Pi-4号（Pi-ta[sup]2[/sup],Pi-sh)与江苏省稻瘟病菌有较高的适合度。品种K3（Pi-k[sup]h[/sup]）在江苏省有较好的抗性,其抗性频率为100%。用两种毒力类型的不同菌株2003-184(ZC[sub]5[/sub])和 2003-14-1(ZG[sub]1[/sub]) 混合接种13个日本已知抗性基因品种后,发现对上述两个单个菌株表现抗病的K3（Pi-k[sup]h[/sup]）品种表现为感病,提示不同毒力的菌株之间的相互作用导致菌株毒力的改变可能是品种抗性丧失的原因之一。     

Occurrence and characterization of fungi and oomycetes transmitted via potting mixtures and organic manures

A study was conducted to investigate the most common fungal and oomycete pathogens introduced into farms in Oman via potting mixtures and organic manures. A total of 37 commercial types of potting mixtures (2 local and 35 imported from overseas), 4 commercial types of organic manures and 11 non-commercial types of organic manures were included in the study. Identification of the isolated species was based on morphological characteristics, except for the most common species which were further identified using sequences of the internal transcribed spacer region of the ribosomal DNA (ITS rDNA). Fusarium spp. (14%), Pythium aphanidermatum (3%), Alternaria spp. (5%), Helminthosporium spp. (5%) and Cladosporium spp. (3%) were recovered at different frequencies from samples of potting mixtures. Fusarium solani (40%) and Fusarium equiseti (47%) were recovered at high frequencies from samples of organic manures. Isolations from organic manures also yielded Pythium periplocum (7%), Rhizoctonia solani (7%), Fusarium lichenicola (7%), Helminthosporium spp. (27%) and Alternaria spp. (27%). Trichoderma spp., Penicillium spp., Aspergillus spp. and Rhizopus spp. were found to be common in samples of potting mixtures and organic manures. Investigating sensitivity to hymexazol among 9 isolates of F. equiseti and 13 isolates of F. solani revealed variations among different isolates. The EC₅₀ values ranged from 1 to over 1200 (avg. 192 μg ml⁻¹) for F. equiseti isolates and from 135 to 789 (avg. 324 μg ml⁻¹) for F. solani isolates, indicating presence of resistance to this important fungicide among some Fusarium isolates. This appears to be the first report of contamination with R. solani, P. periplocum, F. solani, F. equiseti and F. lichenicola of organic manures. This study appears to report for the first time F. lichenicola in Oman and appears to be the first report of occurrence of resistance to hymexazol among F. equiseti and F. solani isolates.     

The Race Structure of the Rice Blast Pathogen Across Southern and Northeastern China

Background

Rice blast, caused by the ascomycete Magnaporthe oryzae (Mo), imposes a major constraint on rice productivity. Managing the disease through the deployment of host resistance requires a close understanding of race structure of the pathogen population.

Results

The host/pathogen interaction between isolates sampled from four Mo populations collected across the rice-producing regions of China was tested using two established panels of differential cultivars. The clearest picture was obtained from the Chinese cultivar panel, for which the frequency of the various races, the race diversity index, the specific race isolate frequency, and the frequency of the three predominant races gave a consistent result, from which it was concluded that the pathogen population present in the southern production region was more diverse than that in the northeastern region. The four blast resistance genes Pi1, Pik, Pik-m, and Piz all still remain effective in the southern China rice production area, as does Pi1 in the northeastern region. The effectiveness of Pita, Pik-p, Piz, and Pib is restricted to single provinces. The distinctive resistance profile shown by the Chinese differential cultivar set implied the presence of at least five as yet unidentified blast resistance genes.

Conclusions

The Chinese differential cultivar set proved to be more informative than the Japanese one for characterizing the race structure of the rice blast pathogen in China. A number of well characterized host resistance genes, in addition to some as yet uncharacterized ones, remain effective across the major rice production regions in China.

     

Sensitivity to iprodione, difenoconazole and fludioxonil of Rhizoctonia cerealis isolates collected from wheat in China

Since the 1980s wheat sharp eyespot (WSE) caused mainly by Rhizoctonia cerealis has become one of serious diseases of wheat in China. In this study, the sensitivity of 89 R. cerealis isolates to different fungicides was evaluated using mycelial growth inhibition assays. The results showed that all R. cerealis isolates tested were sensitive to iprodione, difenoconazole and fludioxonil with mean EC₅₀ (effective concentration that results in 50% of mycelial growth inhibition) values of 0.419, 0.062 and 0.033 μg/ml, respectively. To evaluate the risk of fungicide resistance in R. cerealis, an attempt was made to induce resistant mutants in the laboratory. Although difenoconazole- and fludioxonil-resistant mutants were not obtained, we obtained seven independently iprodione-resistant (IR) mutants from 89 parental isolates. The EC₅₀ values for these IR mutants were greater than 100 μg/ml, whereas those for the original wild-type counterparts were less than 1 μg/ml. After having been subcultured on PDA for 10 generations, the IR mutants did not show any decrease in resistance to iprodione. Additionally, these IR mutants also showed resistance to fludioxonil but remained sensitive to difenoconazole. Osmotic sensitivity tests showed that the IR mutants were hypersensitive to osmotic stress generated by NaCl. Inoculation tests showed that all the IR mutants lost their ability to infect the host plant. Taken together, these results indicate that the current population of R. cerealis is sensitive to these fungicides and a fitness cost is associated with iprodione-resistant mutants of R. cerealis in both osmotic stress and pathogenicity. The information obtained in this study is useful in monitoring and managing fungicide resistance in R. cerealis populations in China.     

Weed competitiveness of the lowland rice varieties of NERICA in the southern Guinea Savanna

Weed competition is a major constraint to lowland rice production in West Africa. Interspecific rice varieties named New Rice for Africa (NERICA) may have superior weed competitiveness and could as such play an important role in integrated weed management. The NERICA varieties were developed from the wide cross between high-yielding Oryza sativa (L.) and weed competitive and disease resilient Oryza glaberrima (Steud.). In this study weed competitiveness of all 60 lowland varieties of NERICA (NERICA-L) was compared with their most frequently used parents [IR64 (O. sativa) and TOG5681 (O. glaberrima)], the weed competitive variety Jaya (O. sativa) and the O. glaberrima upland NERICA parent CG14. During the 2006 and 2007 rainy seasons these varieties were grown under weed-free and weedy conditions in a lowland farmers’ field with partially controlled irrigation in south-east Benin. Weedy plots included single hand weeding at 28 days after sowing, whereas weed-free plots were weekly weeded.     

Molecular Scree ning of Blast Resistance Genes in Rice Germplasms Resistant to Magnaporthe oryzae

Molecular screening of major rice blast resistance genes was determined with molecular markers, which showed close-set linkage to 11 major rice blast resistance genes (Pi-d2, Pi-z, Piz-t, Pi-9, Pi-36, Pi-37, Pi5, Pi-b, Pik-p, Pik-h and Pi-ta²), in a collection of 32 accessions resistant to Magnaporthe oryzae. Out of the 32 accessions, the Pi-d2 and Pi-z appeared to be omnipresent and gave positive express. As the second dominant, Pi-b and Piz-t gene frequencies were 96.9% and 87.5%. And Pik-h and Pik-p gene frequencies were 43.8% and 28.1%, respectively. The molecular marker linkage to Pi-ta² produced positive bands in eleven accessions, while the molecular marker linkage to Pi-36 and Pi-37 in only three and four accessions, respectively. The natural field evaluation analysis showed that 30 of the 32 accessions were resistant, one was moderately resistant and one was susceptible. Infection types were negatively correlated with the genotype scores of Pi-9, Pi5, Pi-b, Pi-ta² and Pik-p, although the correlation coefficients were very little. These results are useful in identification and incorporation of functional resistance genes from these germplasms into elite cultivars through marker-assisted selection for improved blast resistance in China and worldwide.     

Genetic analysis for resistance to the green leafhopper, Nephotettix virescens (Distant), in some cultivars of rice, Oryza sativa L.

The genetics of resistance to green leafhopper Nephotettix virescens (Distant) was studied in 17 cultivars of rice (Oryza sativa L.). Seedlings at the one-leaf stage were infested with second- and third-instar nymphs of green leafhopper. Reactions to seedling injury were recorded 7–8 days after infestation. The reactions of F₁, F₂, and F₃ populations from the crosses of resistant varieties with susceptible parent TN1 revealed that resistance of Tilakkachray, Kalimekri 77-5, and Tilockkachari was governed by two dominant genes and resistance of IR42 was conveyed by a single recessive gene. Single dominant genes govern resistance in the remaining 13 cultivars. Allele tests revealed that a single dominant gene of Segon Perak and one of the two genes of Kalimekri 77-5 are allelic to Glh 1. The single dominant gene of Ernest, Bignou and ARC614 and one of the two genes of each of Tilakkachray, Kalimekri 77-5, and Tilockkachari are allelic to Glh 2. A single dominant gene, which is allelic to Glh 3, conditions resistance in Arai and IR30. The single recessive gene which conveys resistance in IR42 appears to be allelic to glh 4. The allelic relationships of one of the two genes of Tilakkachray and Tilockkachari and single dominant genes of Fulkari 653, Chao Pho Kha, Amoule Borome, Kabero, Aus Murali, IR28 and IR34, with each other and with Glh 5 are not known.     

Effect of plant growth promoting rhizobia on seed germination, growth promotion and suppression of Fusarium wilt of fenugreek (Trigonella foenum-graecum L.)

Five bacterial strains (TR1 to TR5) isolated from root nodules of fenugreek (Trigonella foenum-graecum) were tested for their plant growth promotory traits and biocontrol potential against Fusarium oxysporum. On the basis of morphological, physiological, biochemical and molecular characteristics, strains TR1 and TR3 - TR5 were identified as Ensifer meliloti, and TR2 as Rhizobium leguminosarum. All bacterial isolates utilized phosphate in vitro. Except TR5, all isolates produced IAA and none of them showed volatile cyanogens production. Except TR3, all isolates produced in vitro siderophore. Isolate TR1 and TR4 showed chitinase production while only TR2 showed β-1,3-glucanase activity. Isolates TR1, TR2 and TR5 exhibited ACC deaminase activity. Isolates TR1, TR2 and TR4 inhibited the growth of F. oxysporum, causing loss of structural integrity of the mycelium, hyphal perforation, lysis, fragmentation and degradation. The potential for nodulation and nitrogen fixation of the strains were confirmed by amplification of 500 bp nodC and 781 bp nifH fragments. The application of the TR1 + TR2 combination resulted in increased grain yield by 35% and 36% of fenugreek in two consecutive field trials, respectively as compared to control. Maximum increments in vigour index, nodule number and root and shoot biomass were recorded with seed inoculated with consortium (TR1 + TR2) followed by single inoculation as compared to control. The antibiotic resistant marker strain of E. meliloti TR1^strep+ and R. leguminosarum TR2^tet+ confirmed the efficient colonization of fenugreek roots. This study showed that these rhizobial isolates have properties of biocontrol agents and may be applied to promote the growth of fenugreek.     

Race Specificity of Major Rice Blast Resistance Genes to Magnaporthe grisea Isolates Collected from indica Rice in Guangdong, China

Race-specific resistance and field resistance of 30 rice blast resistance monogenic lines dedved from different resources were evaluated. The spectra of resistance to 163 Magnaporthe grisea isolates collected from indica rice in Guangdong Province, China ranged from 0.6% to 89.6%. Most of the monogenic lines showed a narrow resistance spectrum and high susceptibility in rice blast area, whereas the lines with Pikh and Pi1(t) had the broad resistance spectra of 89.6% and 82.2% respectively, showing a high and stable blast resistance in fields. According to the cluster analysis of specific resistance to 163 blast isolates tested, the 30 monogenic lines were divided into 15 groups, and based on the principal factor analysis, nine kinds of race-specific resistance were identified. Pik, Piz5, Pi9 and Pish can be used as candidate resistance genes for rice breeding since their specific resistance differed from those of the backbone parents in Guangdong, China. Gene pyramiding of Pikh [or Pi1(t)], Pi9 (or Piz5) and Pish (or Pita2) will be effective to obtain broad-spectrum blast resistance in rice breeding program in Guangdong, China. The strategies for studying and application of rice blast resistance genes were discussed.     

Near-isogenic lines of IR64 (Oryza sativa subsp. indica cv.) introgressed with DEEPER ROOTING 1 and STELE TRANSVERSAL AREA 1 improve rice yield formation over the background parent across three water management regimes

Three near-isogenic lines (NILs) of Oryza sativa subsp. indica cv. IR64 (Dro1-NIL, Sta1-NIL, Dro1+Sta1-NIL) with DEEPER ROOTING 1 (DRO1), a novel gene for steeper root growth angle, and/or with Stele Transversal Area 1 (Sta1), a QTL for wider stele area, were tested under flooded lowland (FL), alternate wetting and drying lowland (AWD), and rainfed upland (UP) conditions in 2013 and 2014 to compare the effects of DRO1 and Sta1 on yield across different water management regimes. Genotypic variation and water management effects were significant for grain yield, aboveground biomass, and harvest index, as well as their interactions with year, but no significant genotype × water interaction was detected. Dro1-NIL had 14% higher yield than that of IR64 across the three water conditions due to higher harvest index, aboveground biomass, leaf area index, and number of grains. Sta1 tended to reduce the carbon isotope composition (δ¹³C), leading to a higher harvest index of Sta1-NIL than that of IR64, but grain yield was not increased. Dro1+Sta1-NIL had the highest fraction of intercepted radiation, cumulative radiation interception, and panicle number, with a small but insignificant yield improvement over IR64, but the combination of DRO1 and Sta1 did not surpass the increment from the effects of DRO1 alone. AWD in the more rainy year 2014 attained both higher water productivity and higher biomass, with significant water by year interaction for water productivity. Genotypic variation in water productivity was related with higher leaf area index and fraction interception, with Dro1-NIL larger than in IR64 and Sta1-NIL.     

Effects of genes increasing the number of spikelets per panicle,TAW1 and APO1, on yield and yield-related traits in rice

The genes TAWAWA1 (TAW1) and ABERRANT PANICLE ORGANIZATION1 (APO1) increase the number of spikelets per panicle (SN). In the present study, we examined the effects of these genes on morphological traits, yield, and yield-related traits including yield components using the near-isogenic lines (NILs) in the genetic background of a japonica rice variety, Koshihikari – NIL-taw1, NIL-apo1-D3, and NIL-apo1-D4 – in a field experiment. The SN and total number of spikelets per area of the three NILs were larger than those of Koshihikari. However, the yield of the three NILs did not exceed that of Koshihikari due to their low filling ability. Interestingly, our field experiments indicated that TAW1 did not affect the diameter of internodes and the PN, whereas APO1 decreased the PN and increased the diameter of internodes. These results suggest that TAW1 and APO1 differently affect yield-related traits.