Identification and fine mapping of <Emphasis Type="Italic">qWBPH11</Emphasis> conferring resistance to whitebacked planthopper (<Emphasis Type="Italic">Sogatella furcifera</Emphasis> Horvath) in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

The whitebacked planthopper (WBPH), Sogatella furcifera Horvath, is one of the most destructive pests in rice (Oryza sativa L.) production. Host-plant resistance has been considered as an efficient and eco-friendly strategy to reduce yield losses caused by WBPH. In this study, we found that an indica rice cultivar IR54751-2-44-15-24-2 (IR54751) displayed high resistance to WBPH at both seedling and tillering stages. The resistance of IR54751 was mainly contributed by antixenosis and tolerance rather than antibiosis. An F₂ population derived from a cross between IR54751 and a susceptible japonica cultivar 02428 was constructed to detect the quantitative trait loci (QTLs) conferring the resistance to WBPH. In total, four QTLs including qWBPH3.1, qWBPH3.2, qWBPH11, and qWBPH12 were identified and distributed on three different chromosomes. The four QTLs had LOD scores of 3.8, 8.2, 5.8, and 3.9, accounting for 8.2, 21.5, 13.9, and 10.4% of the phenotypic variation, respectively. Except for qWBPH3.1, the resistance alleles of the other three QTLs were all from IR54751. Further, a secondary population harboring only single qWBPH11 locus was developed from the F₂ population by marker-assisted selection. Finally, qWBPH11 was delimited in a 450-kb region between markers DJ53973 and SNP56. The identification of WBPH resistance QTLs and the fine mapping of qWBPH11 will be helpful for cloning resistance genes and breeding resistant rice cultivars.     

QTL mapping of panicle blast resistance in japonica landrace heikezijing and its application in rice breeding

The rice blast caused by Magnaporthe oryzae is one of the most devastating diseases worldwide, and the panicle blast could result in more loss of yield in rice production. However, the quantitative trait loci (QTLs) and genes related to panicle-blast resistance have not been well studied due to the time-consuming screening methodology involved and variation in symptoms. The QTLs for panicle blast resistance have been mapped in a population of 162 RILs (recombination inbreeding lines), derived from a cross between a highly blast-resistant rice landrace, Heikezijing, and a susceptible variety, Suyunuo. Two QTLs for panicle-blast resistance, qPbh-11–1 and qPbh-7-1, were identified, which were distributed on chromosomes 11 and 7. The QTL qPbh-11–1 was stably detected in three independent experiments, at Nanjing in 2013 and 2014 and at Hainan in 2014, located between the region of RM27187 and RM27381 on the distal end of chromosome 11 far from the reported resistant loci Pb1 and qPbm11 for panicle blast. The QTL qPbh-7-1 was detected only at Nanjing in 2013 and located between the region of M18 and RM3555 on chromosome 7. With marker-assisted selection (MAS) three introgression lines with the major panicle blast-resistance QTL qPbh-11–1 were developed from a recurrent parent Nanjing 44 (NJ44) and the panicle resistance of introgression lines was improved 46.36–55.47 % more than NJ44. Based on the results provided, Heikezijing appears to be a valuable source for panicle blast resistance.     

Genetic and biochemical mechanisms of rice resistance to planthopper

Key message

This article presents a comprehensive review on the genetic and biochemicalmechanisms governing rice-planthopper interactions, aiming to contribute substantialplanthopper control and facilitate breeding for resistance to planthoppers in rice.

Abstract

The rice planthopper is the most destructive pest of rice and a substantial threat to rice production. The brown planthopper (BPH), white-backed planthopper (WBPH) and small brown planthopper (SBPH) are three species of delphacid planthoppers and important piercing-sucking pests of rice. Host-plant resistance has been recognized as the most practical, economical and environmentally friendly strategy to control planthoppers. Until now, at least 30, 14 and 34 major genes/quantitative trait loci for resistance to BPH, WBPH and SBPH have been identified, respectively. Recent inheritance and molecular mapping of gene analysis showed that some planthopper-resistance genes in rice derived from different donors aggregate in clusters, while resistance to these three species of planthoppers in a single donor is governed not by any one dominant gene but by multiple genes. Notably, Bph14, Bph26, Bph3 and Bph29 were successfully identified as BPH-resistance genes in rice. Biological and chemical studies on the feeding of planthoppers indicate that rice plants have acquired various forms of defence against planthoppers. Between the rice-planthopper interactions, rice plants defend against planthoppers through activation the salicylic acid-dependent systemic acquired resistance but not jasmonate-dependent hormone response pathways. Transgenic rice for the planthopper-resistance mechanism shows that jasmonate and its metabolites function diversely in rice’s resistance to planthopper. Understanding the genetic and biochemical mechanisms underlying resistance in rice will contribute to the substantial control of such pests and facilitate breeding for rice’s resistance to planthopper more efficiently.

     

Locating QTLs controlling overwintering seedling rate in perennial glutinous rice 89-1 (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

A new cold tolerant germplasm resource named glutinous rice 89-1 (Gr89-1, Oryza sativa L.) can overwinter using axillary buds, with these buds being ratooned the following year. The overwintering seedling rate (OSR) is an important factor for evaluating cold tolerance. Many quantitative trait loci (QTLs) controlling cold tolerance at different growth stages in rice have been identified, with some of these QTLs being successfully cloned. However, no QTLs conferring to the OSR trait have been located in the perennial O. sativa L. To identify QTLs associated with OSR and to evaluate cold tolerance. 286 F₁₂ recombinant inbred lines (RILs) derived from a cross between the cold tolerant variety Gr89-1 and cold sensitive variety Shuhui527 (SH527) were used. A total of 198 polymorphic simple sequence repeat (SSR) markers that were distributed uniformly on 12 chromosomes were used to construct the linkage map. The gene ontology (GO) annotation of the major QTL was performed through the rice genome annotation project system. Three main-effect QTLs (qOSR2, qOSR3, and qOSR8) were detected and mapped on chromosomes 2, 3, and 8, respectively. These QTLs were located in the interval of RM14208 (35,160,202 base pairs (bp))–RM208 (35,520,147 bp), RM218 (8,375,236 bp)–RM232 (9,755,778 bp), and RM5891 (24,626,930 bp)–RM23608 (25,355,519 bp), and explained 19.6%, 9.3%, and 11.8% of the phenotypic variations, respectively. The qOSR2 QTL displayed the largest effect, with a logarithm of odds score (LOD) of 5.5. A total of 47 candidate genes on the qOSR2 locus were associated with 219 GO terms. Among these candidate genes, 11 were related to cell membrane, 7 were associated with cold stress, and 3 were involved in response to stress and biotic stimulus. OsPIP1;3 was the only one candidate gene related to stress, biotic stimulus, cold stress, and encoding a cell membrane protein. After QTL mapping, a total of three main-effect QTLs—qOSR2, qOSR3, and qOSR8—were detected on chromosomes 2, 3, and 8, respectively. Among these, qOSR2 explained the highest phenotypic variance. All the QTLs elite traits come from the cold resistance parent Gr89-1. OsPIP1;3 might be a candidate gene of qOSR2.     

Genetic mapping and confirmation of quantitative trait loci for grain chalkiness in rice

Grain chalkiness is a highly undesirable trait affecting rice grain quality and milled rice yield. In order to clarify the genetic basis of chalkiness, a recombinant inbred line population (RIL) derived from a cross between Beilu130 (a japonica cultivar with high chalkiness) and Jin23B (an indica cultivar with low chalkiness) was developed for quantitative trait locus (QTL) mapping. A total of 10 QTLs for white belly rate (WBR) and white core rate (WCR) were detected on eight different chromosomes over 2 years. Two QTLs for WBR (qWBR2 and qWBR5) showed similar chromosomal locations with GW2 and qSW5/GW5, which have been cloned previously to control the grain width and should be the right candidate genes. Three novel minor QTLs controlling WCR, qWCR1, qWCR3, and qWCR4 were further validated in near isogenic F₂ populations (NIL-F₂) and explained 26.1, 18.3, and 21.1% of the phenotypic variation, respectively. These QTLs could be targets for map-based cloning of the candidate genes to elucidate the molecular mechanism of chalkiness and for marker-assisted selection (MAS) in rice grain quality improvement.     

Identification of Novel Strain-Specific and Environment-Dependent Minor QTLs Linked to Fire Blight Resistance in Apples

Since its first report almost 200 years ago, fire blight, caused by the gram-negative bacterium Erwinia amylovora, has threatened apple and pear production globally. Identifying novel genes and their functional alleles is a prerequisite to developing apple cultivars with enhanced fire blight resistance. Here, we report 13 strain-specific and environment-dependent minor QTLs linked to fire blight resistance from a segregating Malus sieversii × Malus × domestica mapping population. Interval mapping at 95% confidence and Kruskal–Wallis analysis at P value =?0.005 were used to identify QTLs for three strains of E. amylovora differing in virulence and pathogenicity. The QTLs identified explain a small to moderate part of resistance variability, and a majority was not common between years or E. amylovora strains. These QTLs are distributed in eight linkage groups of apples and comparison of their map position to previously identified fire blight resistance QTLs indicates that most are novel loci. Interaction between experimental conditions in the greenhouse and field, and between years, and differences in virulence levels of strains might be responsible for strain- and year-specific QTLs. The QTLs identified on LG10 for strain Ea273 in 2011 and strain LP101 in 2011, and on LG15 for strain LP101 could be the same QTLs identified previously with strain CFBP1430 in cultivar “Florina” and “Co-op16 × Co-op17” mapping population, respectively. We discuss the potential impact of newly identified minor fire blight QTLs and major gene-based resistance on the rate of mutation in pathogen populations to overcome resistance and durability of resistance.     

QTLs for heading date and plant height under multiple environments in rice

Both heading date and plant height are important traits related to grain yield in rice. In this study, a recombinant inbred lines (RILs) population was used to map quantitative trait loci (QTLs) for both traits under 3 long-day (LD) environments and 1 short-day (SD) environment. A total of eight QTLs for heading date and three QTLs for plant height were detected by composite interval mapping under LD conditions. Additional one QTL for heading date and three QTLs for plant height were identified by Two-QTL model under LD conditions. Among them, major QTLs qHd7.1, qHd7.2 and qHd8 for heading date, and qPh1 and qPh7.1 for plant height were commonly detected. qHd7.1 and qHd7.2 were mapped to small regions of less than 1 cM. Genome position comparison of previously cloned genes with QTLs detected in this study revealed that qHd5 and qPh3.1 were two novel QTLs. The alleles of these QTLs increasing trait values were dispersed in both parents, which well explained the transgressive segregation observed in this population. In addition, the interaction between qHd7.1 and qHd8 was detected under all LD conditions. Multiple-QTL model analysis revealed that all QTLs and their interactions explained over 80% of heading date variation and 50% of plant height variation. Two heading date QTLs were detected under SD condition. Of them, qHd10 were commonly identified under LD condition. The difference in QTL detection between LD and SD conditions indicated most heading date QTLs are sensitive to photoperiod. These findings will benefit breeding design for heading date and plant height in rice.     

Combining drought and submergence tolerance in rice: marker-assisted breeding and QTL combination effects

TDK1 is a popular rice variety from the Lao PDR. Originally developed for irrigated conditions, this variety suffers a high decline in yield under drought conditions. Studies have identified three quantitative trait loci (QTLs) for grain yield under drought conditions, qDTY _3.1 , qDTY _6.1 , and qDTY _6.2 , that show a high effect in the background of this variety. We report here the pyramiding of these three QTLs with SUB1 that provides 2–3 weeks of tolerance to complete submergence, with the aim to develop drought- and submergence-tolerant near-isogenic lines (NILs) of TDK1. We used a tandem approach that combined marker-assisted backcross breeding with phenotypic selection to develop NILs with high yield under drought stress and non-stress conditions and preferred grain quality. The effect of different QTL combinations on yield and yield-related traits under drought stress and non-stress conditions is also reported. Our results show qDTY _3.1 to be the largest and most consistent QTL affecting yield under drought conditions, followed by qDTY _6.1 and qDTY _6.2 , respectively. QTL class analysis also showed that lines with a combination of qDTY _3.1 and qDTY _6.1 consistently showed a higher tolerance to drought than those in which one of these QTLs was missing. In countries such as Lao PDR, where large areas under rice cultivation suffer vegetative-stage submergence and reproductive-stage drought, these lines could ensure yield stability. These lines can also serve as valuable genetic material to be used for further breeding of high-yielding, drought- and submergence-tolerant varieties in local breeding programs.     

Genetic analysis and mapping of adult plant resistance loci to leaf rust in durum wheat cultivar Bairds

Key message

New leaf rust adult plant resistance (APR) QTL QLr.cim - 6BL was mapped and confirmed the known pleotropic APR gene Lr46 effect on leaf rust in durum wheat line Bairds.

Abstract

CIMMYT-derived durum wheat line Bairds displays an adequate level of adult plant resistance (APR) to leaf rust in Mexican field environments. A recombinant inbred line (RIL) population developed from a cross of Bairds with susceptible parent Atred#1 was phenotyped for leaf rust response at Ciudad Obregon, Mexico, during 2013, 2014, 2015 and 2016 under artificially created epidemics of Puccinia triticina (Pt) race BBG/BP. The RIL population and its parents were genotyped with the 50 K diversity arrays technology (DArT) sequence system and simple sequence repeat (SSR) markers. A genetic map comprising 1150 markers was used to map the resistance loci. Four significant quantitative trait loci (QTLs) were detected on chromosomes 1BL, 2BC (centromere region), 5BL and 6BL. These QTLs, named Lr46, QLr.cim-2BC, QLr.cim-5BL and QLr.cim-6BL, respectively, explained 13.5–60.8%, 9.0–14.3%, 2.8–13.9%, and 11.6–29.4%, respectively, of leaf rust severity variation by the inclusive composite interval mapping method. All of these resistance loci were contributed by the resistant parent Bairds, except for QLr.cim-2BC, which came from susceptible parent Atred#1. Among these, the QTL on chromosome 1BL was the known pleiotropic APR gene Lr46, whereas QLr.cim-6BL, a consistently detected locus, should be a new leaf rust resistance locus in durum wheat. The mean leaf rust severity of RILs carrying all four QTLs ranged from 8.0 to 17.5%, whereas it ranged from 10.9 to 38.5% for three QTLs (Lr46 + 5BL + 6BL) derived from the resistant parent Bairds. Two RILs with four QTLs combinations can be used as sources of complex APR in durum wheat breeding.

     

Mapping of QTLs associated with resistance to common bunt,tan spot,leaf rust,and stripe rust in a spring wheat population

Spring wheat (Triticum aestivum L.) breeding goals in western Canada include good agronomic characteristics and good end-use quality, and also moderate to elevated resistance to diseases of economic importance. In this study, we aimed to identify quantitative trait loci (QTL) associated with resistance to common bunt (Tilletia tritici and Tilletia laevis), tan spot (Pyrenophora tritici-repentis), leaf rust (Puccinia triticina), and stripe rust (Puccinia striiformis f. sp. tritici). A total of 167 recombinant inbred lines (RILs) derived from a cross between two spring wheat cultivars, ‘Attila’ and ‘CDC Go’, were evaluated for reactions to the four diseases in nurseries from three to eight environments, and genotyped with the Wheat 90K SNP array and three gene-specific markers (Ppd-D1, Vrn-A1, and Rht-B1). The RILs exhibited transgressive segregation for all four diseases, and we observed several lines either superior or inferior to the parents. Broad-sense heritability varied from 0.25 for leaf rust to 0.48 for common bunt. Using a subset of 1203 informative markers (1200 SNPs and 3 gene-specific markers) and average disease scores across all environments, we identified two QTLs (QCbt.dms-1B.2 and QCbt.dms-3A) for common bunt, and three QTLs each for tan spot (QTs.dms-2B, QTs.dms-2D, and QTs.dms-6B), leaf rust (QLr.dms-2D.1, QLr.dms-2D.2, and QLr.dms-3A), and stripe rust (QYr.dms-3A, QYr.dms-4A, and QYr.dms-5B). Each QTL individually explained between 5.9 and 18.7% of the phenotypic variation, and altogether explained from 21.5 to 26.5% of phenotypic and from 52.2 to 86.0% of the genetic variation. The resistance alleles for all QTLs except one for stripe rust (QYr.dms-5B) were from CDC Go. Some of the QTLs are novel, while others mapped close to QTLs and/or genes reported in other studies.     

Marker-aided selection and validation of various $${ }$$ gene combinations for rice blast resistance in elite rice variety ADT 43

Rice blast caused by fungal pathogen Pyricularia oryzae has a major impact on reducing yield potential of rice. In this study, homozygous plants were selected using microsatellite markers from the \(\hbox {BC}_{3}\hbox {F}_{2}\) population pyramided with four major genes in elite rice variety ADT 43. Background and selected lines with various blast resistance gene combinations were screened under natural conditions to study the effects of various gene combinations. Upon inspection of lines with different gene combinations, the three-gene pyramided line Pi54+Pi33+Pi1 was found to be highly resistant with the score of 3.3 followed by other three-gene pyramided lines Pi54+Pi2+Pi1 and Pi33+Pi2+Pi1, with the scores of 3.9 and 3.8, respectively. Two-gene pyramided lines Pi54+Pi1, Pi33+Pi1 and Pi2+Pi1 were found to be moderately resistant with a mean score of 4.0 each. In the case of monogenic lines, positive plants for Pi54 performed almost equal to three-gene pyramided lines with a mean score of 3.6. Lines with Pi2 and Pi1 were found to be moderately resistant and moderately susceptible with the mean scores of 4.1 and 4.5, respectively.     

Evaluation of QTLs for Shoot Fly (<Emphasis Type="Italic">Atherigona soccata</Emphasis>) Resistance Component Traits of Seedling Leaf Blade Glossiness and Trichome Density on Sorghum (<Emphasis Type="Italic">Sorghum bicolor</Emphasis>) Chromosome SBI-10L

Shoot fly is a major insect pest of sorghum damaging early crop growth, establishment and productivity. Host plant resistance is an efficient approach to minimize yield losses due to shoot fly infestation. Seedling leaf blade glossiness and trichome density are morphological traits associated with shoot fly resistance. Our objective was to identify and evaluate QTLs for glossiness and trichome density using- i) 1894 F₂s, ii) a sub-set of 369 F₂-recombinants, and iii) their derived 369 F_2:3 progenies, from a cross involving introgression lines RSG04008-6 (susceptible)?×?J2614-11 (resistant). The QTLs were mapped to a 37–72 centimorgan (cM) or 5–15 Mb interval on the long arm of sorghum chromosome 10 (SBI-10L) with flanking markers Xgap001 and Xtxp141. One QTL each for glossiness (QGls10) and trichome density (QTd10) were mapped in marker interval Xgap001-Xnhsbm1044 and Xisep0630-Xtxp141, confirming their loose linkage, for which phenotypic variation accounted for ranged from 2.29 to 11.37 % and LOD values ranged from 2.03 to 24.13, respectively. Average physical map positions for glossiness and trichome density QTLs on SBI-10 from earlier studies were 4 and 2 Mb, which in the present study were reduced to 2 Mb and 800 kb, respectively. Candidate genes Glossy15 (Sb10g025053) and ethylene zinc finger protein (Sb10g027550) falling in support intervals for glossiness and trichome density QTLs, respectively, are discussed. Also we identified a sub-set of recombinant population that will facilitate further fine mapping of the leaf blade glossiness and trichome density QTLs on SBI-10.     

Marker-assisted pyramiding of bacterial blight and gall midge resistance genes into RPHR-1005, the restorer line of the popular rice hybrid DRRH-3

Bacterial blight (BB) of rice caused by the pathogen Xanthomonas oryzae pv. oryzae and the insect gall midge (GM) (Orseolia oryzae) are two major constraints of rice production. The present study was carried out to improve RPHR-1005, a stable restorer line of the fine-grain-type rice hybrid DRRH-3, for BB and GM resistance through marker-assisted backcross breeding (MABB). Two major GM resistance genes, Gm4 and Gm8, and a major BB resistance gene, Xa21, were selected as target genes for transfer to RPHR-1005. Two sets of backcrosses were carried out to combine either Xa21 + Gm4 or Xa21+ Gm8 into RPHR-1005 using breeding lines in the genetic background of ISM possessing either Gm4 or Gm8 along with Xa21. Foreground selection was performed for Xa21, Gm4, Gm8, and the major fertility restorer genes Rf3 and Rf4 using gene-specific markers, while 61 polymorphic simple sequence repeat (SSR) markers were used for background selection and marker-assisted backcrossing was continued until BC₂ generation. A promising homozygous backcross-derived plant at the BC₂F₂ generation possessing Xa21 + Gm4, and another possessing Xa21 + Gm8, were intercrossed to stack the target resistance genes. At ICF ₄ (inter-crossed F₄) , three promising lines possessing the three target resistance genes in a homozygous condition along with fine-grain type, complete fertility restoration, and better panicle exsertion than RPHR-1005 have been identified. Among these, a single line, # RPIC-16-65-125, showed better yield, was highly resistant to BB and GM, was of medium–slender grain type, and had complete fertility restoration along with better panicle exsertion and taller plant type than RPHR-1005. This is the first report of combining resistance against BB and GM in the genetic background of a hybrid rice parental line.     

Temperature-dependent QTLs in <Emphasis Type="Italic">indica</Emphasis> alleles for improving grain quality in rice: increased prominence of QTLs responsible for reduced chalkiness under high-temperature conditions

Quantitative trait loci (QTLs) for the apparent quality of brown rice under high temperatures during ripening were analyzed using chromosomal segment substitution lines. Segments from the indica cultivar Habataki were substituted into a japonica cultivar with a Sasanishiki background. We found the following two QTLs for increasing grain quality in the Habataki allele on chromosome 3: (1) qTW3-2, located near the marker RM14702, decreased the percentage of total white immature (TWI) grains, and (2) qRG3-2, located near RM3766, increased the percentage of regular grains. The effects of these two QTLs were more obvious under high-temperature ripening conditions; hence, these loci are considered QTLs not only for reducing TWI grains but also for increasing high-temperature tolerance. Additionally, we found two QTLs, i.e., qTW3-1 and qRG3-1, responsible for reduced grain quality near RM14314 on chromosome 3. Although the QTL for narrow grains in the Habataki allele qNG3 was genetically linked to qTW3-2, the effect was only slightly significant, and the length/width ratio of qNG3-carrying grains was within the range observed in widely grown japonica cultivars. Incorporating the Habataki region, including qRG3-2 and qTW3-2 but not qTW3-1 and qRG3-1, in addition to previously reported grain quality QTLs in breeding japonica cultivars will improve high-temperature tolerance and grain quality.     

QTL mapping for downy mildew resistance in cucumber via bulked segregant analysis using next-generation sequencing and conventional methods

Key message

QTL mapping using NGS-assisted BSA was successfully applied to an F ₂ population for downy mildew resistance in cucumber. QTLs detected by NGS-assisted BSA were confirmed by conventional QTL analysis.

Abstract

Downy mildew (DM), caused by Pseudoperonospora cubensis, is one of the most destructive foliar diseases in cucumber. QTL mapping is a fundamental approach for understanding the genetic inheritance of DM resistance in cucumber. Recently, many studies have reported that a combination of bulked segregant analysis (BSA) and next-generation sequencing (NGS) can be a rapid and cost-effective way of mapping QTLs. In this study, we applied NGS-assisted BSA to QTL mapping of DM resistance in cucumber and confirmed the results by conventional QTL analysis. By sequencing two DNA pools each consisting of ten individuals showing high resistance and susceptibility to DM from a F₂ population, we identified single nucleotide polymorphisms (SNPs) between the two pools. We employed a statistical method for QTL mapping based on these SNPs. Five QTLs, dm2.2, dm4.1, dm5.1, dm5.2, and dm6.1, were detected and dm2.2 showed the largest effect on DM resistance. Conventional QTL analysis using the F₂ confirmed dm2.2 (R ² = 10.8–24 %) and dm5.2 (R ² = 14–27.2 %) as major QTLs and dm4.1 (R ² = 8 %) as two minor QTLs, but could not detect dm5.1 and dm6.1. A new QTL on chromosome 2, dm2.1 (R ² = 28.2 %) was detected by the conventional QTL method using an F₃ population. This study demonstrated the effectiveness of NGS-assisted BSA for mapping QTLs conferring DM resistance in cucumber and revealed the unique genetic inheritance of DM resistance in this population through two distinct major QTLs on chromosome 2 that mainly harbor DM resistance.

     

Comparison of rice flowering-time genes under paddy conditions

Heading date is one of most important agronomic traits in rice. Flowering regulatory mechanisms have been elucidated in many cultivars through various approaches. Although study about flowering has been extensively examined in rice, but contributions of floral regulators had been poorly understood in a common genetic background for rice grown under paddy conditions. Thus, we compared the expression of 10 flowering-time genes — OsMADS50, OsMADS51, OsVIL2, OsPhyA, OsPhyB, OsPhyC, Ghd7, Hd1, OsGI, and OsTrx1 — in the same genetic background for ‘Dongjin’ rice (Oryza sativa) grown under paddy conditions when days were longer than 13.5 h. Whereas the wild type (WT) rice flowered 105 days after sowing, the latest mutant to do so was ostrx1, flowering 53 d later. This indicated that the gene is the strongest inducer among all of those examined. Mutations in OsMADS50 delayed flowering by 45 d when compared with the WT, suggesting that this MADS gene is another strong positive element. The third positive element was OsVIL2; mutations in the gene caused plants to flower 27 d late. In contrast, the double phytochrome mutant osphyA osphyB flowered 44 d earlier than the WT. The single mutant osphyB and the double mutant osphyB osphyC did the same, although not as early as the osphyA osphyB double mutant. These results demonstrated that phytochromes are major inhibitors under paddy conditions. Mutations in Ghd7 accelerated flowering by 34 d, indicating that the gene is also a major inhibitor. The hd1 mutants flowered 16 d earlier than the WT while a mutation in OsGI hastened flowering by 10 d, suggesting that both are weak flowering repressors. Of the two florigen genes (Hd3a being the other one), RFT1 played a major role under paddy conditions. Its expression was strongly promoted by Ehd1, which was negatively controlled by Ghd7. Here we show that phytochromes strongly inhibit flowering and OsTrx1 and OsMADS50 significantly induce flowering under paddy conditions through Ghd7-Ehd1-RFT1 pathway. Thus, we may be able to control heading date under paddy conditions through manipulating those genes, Ghd7, Ehd1 and RFT1.     

Improvement of seedling and panicle blast resistance in <Emphasis Type="Italic">Xian</Emphasis> rice varieties following <Emphasis Type="Italic">Pish</Emphasis> introgression

Rice blast is a serious disease caused by the filamentous ascomycetous fungus Magnaporthe oryzae. Incorporating disease resistance genes in rice varieties and characterizing the distribution of M. oryzae isolates form the foundation for enhancing rice blast resistance. In this study, the blast resistance gene Pish was observed to be differentially distributed in the genomes of rice sub-species. Specifically, Pish was present in 80.5% of Geng varieties, but in only 2.3% of Xian varieties. Moreover, Pish conferred resistance against only 23.5% of the M. oryzae isolates from the Geng-planting regions, but against up to 63.2% of the isolates from the Xian-planting regions. Thus, Pish may be an elite resistance gene for improving rice blast resistance in Xian varieties. Therefore, near-isogenic lines (NILs) with Pish and the polygene pyramid lines (PPLs) PPL^Pish/Pi1, PPL^Pish/Pi54, and PPL^Pish/Pi33 in the Xian background Yangdao 6 were generated using a molecular marker-assisted selection method. The results suggested that (1) Pish significantly improved rice blast resistance in Xian varieties, which exhibited considerably improved seedling and panicle blast resistance after Pish was introduced; (2) PPLs with Pish were more effective than the NILs with Pish regarding seedling and panicle blast resistance; (3) the PPL seedling and panicle blast resistance was improved by the complementary and overlapping effects of different resistance genes; and (4) the stability of NIL and PPL resistance varied under different environmental conditions, with only PPL^Pish/Pi54 exhibiting highly stable resistance in three natural disease nurseries (Jianyang, Jinggangshan, and Huangshan). This study provides new blast resistance germplasm resources and describes a novel molecular strategy for enhancing rice blast resistance.     

The diversity of culturable yeasts in the phylloplane of rice in Thailand

One hundred and fifty-six yeast strains were obtained by the enrichment technique from the phylloplanes of 85 rice leaf samples collected from seven provinces in Thailand. On the basis of the D1/D2 domain of the large subunit rRNA gene sequence analysis, 156 strains were identified as 34 known species in 18 genera consisting of 25 species in 13 genera of the phylum Ascomycota and nine species in five genera of the phylum Basidiomycota. The species in the phylum Ascomycota comprised 24 species in 12 genera of the order Saccharomycetales and one species viz. Yarrowia lipolytica in Saccharomycetales incertae sedis. The 24 species viz. Candida glabrata in the Nakaseomyces clade of Saccharomycetaceae, Candida jaroonii, Candida membranifaciens and Candida terebra in the Yamadazyma clade of Debaryomycetaceae, Candida pseudolambica in the Pichia clade of Pichiaceae, Candida ruelliae in the Metschnikowia clade of Metschnikowiaceae, and three unaffiliated clade Candida species (Candida catenulata, Candida rugosa and Candida tropicalis); Clavispora lusitaniae, Kodamaea ohmeri, Metschnikowia koreensis and Metschnikowia lopburiensis in Metschnikowiaceae; Cyberlindnera fabianii, Cyberlindnera rhodanensis and Wickerhamomyces ciferrii in Wickerhamomycetaceae; Debaryomyces nepalensis, Meyerozyma caribbica, Meyerozyma guilliermondii, Millerozyma koratensis, and Yamadazyma mexicanum in Debaryomycetaceae; Pichia kudriavzevii in Pichiaceae; and Lachancea thermotolerans in Saccharomycetaceae. The species in Basidiomycota viz. Cryptococcus flavescens, Cryptococcus laurentii, Cryptococcus aff. laurentii and Cryptococcus rajasthanensis in the Tremellales lineage, Bulleromyces clade, Tremellales, Tremellomycetes, Agaricomycotina; Pseudozyma antarctica and Pseudozyma aphidis in Ustilaginales, Ustilaginomycetes, Ustilaginomycotina; Rhodotorula taiwanensis and Sporobolomyces blumeae in Sporidiobolales, Microbotryomycetes, Pucciniomycotina; and Trichosporon asahii in Trichosporonales, Tremellomycetes, Agaricomycotina. The most prevalent species was R. taiwanensis with a 23 % frequency of occurrence followed by Candida tropicalis (16 %) and Cryptococcus fabianii (12 %).     

Development of near-isogenic lines with different alleles of Piz locus and analysis of their breeding effect under Yangdao 6 background

Rice blast is one of the most destructive diseases of rice. The most effective way of managing this disease is to develop resistant cultivars by introducing resistance genes into elite rice recipients. In this study, the near-isogenic lines (NILs) of six resistance alleles of the Piz locus (Pizt, Pi2, Pigm, Pi40, Pi9 and Piz) were constructed with Yangdao 6 as genetic background. Seedling inoculation tests showed that most of the NILs, namely NIL-Pi2, NIL-Pigm, NIL-Pi9, NIL-Pizt and NIL-Pi40, exhibited good resistance to blast with resistance frequencies (RFs) of over 82.50 %, execpt NIL-Piz which showed lower resistance with a RF of only 36.13 %. Furthermore, the improved-resistance NILs exhibited high similarity of their resistance spectra, with overlapping degrees of resistance spectrum (OD) of more than 75.83 %. However, the RF of panicle blast for all NILs decreased significantly compared with seedling blast in an artificial inoculation test. Although NIL-Pigm showed a higher panicle blast RF of 80 %, other NILs with outstanding performance in seedling blast resistance, namely NIL-Pizt, NIL-Pi2, NIL-Pi9 and NIL-Pi40, exhibited middle or low RFs of panicle blast with values from 56.67 to 33.30 %. Natural induction in a disease nursery showed a consistent trend in artificial inoculation results at seedling and heading stages. While NIL-Pigm was found to exhibit good resistance to leaf blast and panicle blast, NIL-Pi9 and NIL-Pizt were further demonstrated to show excellent resistance in Suichuan, Jiangxi province and Enshi, Hubei province, respectively, because of the race–region specificity. Agronomic traits of NILs were also investigated in order to evaluate the linkage drag effect of different alleles of the Piz locus. The resistance effects of the different alleles of the Piz locus under identical genetic background against seedling blast and panicle blast was first reported in this study, and the above results are expected to provide a theoretical support for the rational utilization of broad-spectrum resistance genes in breeding practice.