A new locus for resistance to brown planthopper identified in the indica rice variety DV85

The brown planthopper (BPH) is one of the most destructive insect pests of rice. Resistant varieties have proved to be one of the most economic and effective measures for BPH management. In this study, an indica rice ‘DV85’ showed resistance to biotype 2 of BPH by bulked seedling test, and a recombinant inbred line (RIL) population derived from a cross between a susceptible rice ‘Kinmaze’ and ‘DV85’ was phenotyped to map genetic factors conferring BPH resistance in ‘DV85′. Composite interval mapping revealed that one quantitative trait locus (QTL) with a LOD score of 10.1 was detected between XNpb202 and C1172 on chromosome 11. This QTL was designated as Qbph11. Qbph11 explained 68.4% of the phenotypic variance of BPH resistance in this population. The allele from the resistant parent ‘DV85’ at Qbph11 reduced the damage caused by BPH feeding and would be very useful in breeding resistant rice varieties via marker‐assisted selection.     

QTL analysis for the resistance to small brown planthopper (Laodelphax striatellus Fallén) in rice using backcross inbred lines

Small brown planthopper (SBPH) is a serious pest of rice ( Oryza sativa L.) in China. An indica variety 'Kasalath' is highly resistant to SBPH. A mapping population consisting of 98 BC₁F₉ lines, derived from a backcross of 'Nipponbare'/'Kasalath'//'Nipponbare', was applied to detect quantitative trait loci (QTL) for resistance to SBPH. In the modified seedbox screening test, three QTLs for SBPH resistance were mapped on chromosomes 3 and 11, explaining 49.9% of the phenotypic variance. In the antixenosis test, a total of three QTLs conferring antixenosis against SBPH were detected on chromosomes 3, 8 and 11, which accounted for 36.4% of the total phenotypic variance. In addition, two QTLs expressing antibiosis to SBPH were detected on chromosomes 2 and 11, explaining 13.8% and 14.7% of the phenotypic variance, respectively. Qsbph11e , Qsbph11f and Qsbph11g were located in the region between S2260 and G257 on chromosome 11, indicating that the locus is significant in conferring resistance to SBPH in 'Kasalath'. The molecular markers linked to these QTLs should be useful in the development of varieties with horizontal resistance to SBPH.     

Multiplex PCR genotyping for five bacterial blight resistance genes applied to marker‐assisted selection in rice (Oryza sativa)

Bacterial blight (BB) is the most economically damaging disease of rice in Asia and other parts of the world. In this study, a multiplex PCR genotyping method was developed to simultaneously identify genotypes of five BB resistance genes, Xa4, xa5, Xa7, xa13 and Xa21. The resistance R alleles were amplified using five functional markers (FMs) to generate amplicons of 217, 103, 179, 381 and 595 bp in IRBB66. Amplicons of 198, 107, 87, 391 and 467 bp corresponded to susceptible alleles in Taiwanese japonica rice cultivars. In backcross breeding programmes, the multiplex PCR assay was integrated into selection from a population using BB resistance donor IRBB66 crossed to rice cultivar ‘Tainung82’. Two plants with homozygosity for Xa4, xa5, Xa7, xa13 and Xa21 were selected from 1100 BC₂F₂ plants. In addition, the five BB resistance genes were also accurately identified in F₂ populations. This multiplex PCR method provides a rapid and efficient method for detecting various BB resistance genes, which will assist in pyramiding genes to improve durability of BB resistance in Taiwanese elite rice cultivars.     

Marker‐assisted introgression of bacterial blight and blast resistance into IR 58025B,an elite maintainer line of rice

IR 58025A is a very popular wild‐abortive cytoplasmic male sterile (WA‐CMS) line of rice and is extensively used for hybrid rice breeding. However, IR 58025A and many hybrids derived from it possess mild aroma (undesirable in some parts of India) and are highly susceptible to bacterial blight (BB) and blast diseases. To improve IR 58025A for BB and blast resistance, we have introgressed a major dominant gene conferring resistance against BB (i.e. Xa21) and blast (i.e. Pi54) into IR 58025B, the maintainer line of IR 58025A. An introgression line of Samba Mahsuri (i.e. SM2154) possessing Xa21 and Pi54 genes in homozygous condition and fine‐grain type was used as donor parent, and backcross breeding strategy was adopted for targeted introgression of the resistance genes. PCR‐based molecular markers tightly linked to Xa21 and Pi54 were used for selection of BB‐ and blast‐resistant lines, while closely linked markers were used for identification of backcross‐derived plants devoid of Rf4 and aroma. At BC₂F₅, four backcross‐derived lines possessing resistance against BB and blast, devoid of aroma, high yield, short plant stature, long‐slender grain type and with recurrent parent genome recovery ranging from 88.8% to 98.6% were selected and advanced for further evaluation. The improved versions of IR 58025B, viz. SB54‐11‐143‐9‐44‐5, SB54‐11‐143‐9‐44‐98, SB54‐11‐143‐9‐44‐111 and SB54‐11‐143‐9‐44‐171, behaved as perfect maintainers when test‐crossed with WA‐CMS lines. Agronomically superior lines of improved IR 58025B are being converted to CMS line through backcrossing for developing high‐yielding and biotic stress‐resistant rice hybrids.     

Construction of high‐throughput genotyped chromosome segment substitution lines in rice (Oryza sativa L.) and QTL mapping for heading date

Chromosome segment substitution lines (CSSLs) provide ideal materials for quantitative trait loci (QTLs) mapping and genetic dissection of complex traits. In this study, we developed a set of CSSL population consisting of 175 lines, which were derived between the recipient ‘Guangluai 4’ and the donor ‘Nipponbare’. Based on 260 molecular markers, we firstly constructed a physical map of core 97 lines. Then, these 97 lines were further genotyped based on resequencing data, and a resequencing‐based physical map was constructed. Compared with the molecular marker‐based physical map, the resequencing‐based physical map of 97 lines contained 367 substituted segments with 252 newly discovered segments. The total size of the 367 substituted segments was 1,074 Mb, which was 2.81 times the size of rice genome. Using the 97 CSSLs as materials, we identified nine QTLs for heading date and three of them were firstly reported. All the QTLs had positive additive effects, ranging from 9.50 to 16.50 days. These CSSLs may greatly help forge a new resource for functional genomics studies and molecular breeding in rice.     

Substitution mapping and characterization of brown planthopper resistance genes from indica rice variety, ‘PTB33’ (Oryza sativa L.)

Rice (Oryza sativa L.) yield is severely reduced by the brown planthopper (BPH), Nilaparvata lugens Stål, in Asian countries. Increasing resistance in rice against BPH can mitigate yield loss. Previous reports indicated the presence of three BPH resistance genes, BPH2, BPH17-ptb, and BPH32, in durable resistant indica rice cultivar ‘PTB33’. However, several important questions remain unclear; the genetic locations of BPH resistance genes on rice chromosomes and how these genes confer resistance, especially with relationship to three major categories of resistance mechanisms; antibiosis, antixenosis or tolerance. In this study, locations of BPH2, BPH17-ptb, and BPH32 were delimited using chromosome segment substitution lines derived from crosses between ‘Taichung 65’ and near-isogenic lines for BPH2 (BPH2-NIL), BPH17-ptb (BPH17-ptb-NIL), and BPH32 (BPH32-NIL). BPH2 was delimited as approximately 247.5 kbp between RM28449 and ID-161-2 on chromosome 12. BPH17-ptb and BPH32 were located between RM1305 and RM6156 on chromosome 4 and RM508 and RM19341 on chromosome 6, respectively. The antibiosis, antixenosis, and tolerance were estimated by several tests using BPH2-NIL, BPH17-ptb-NIL, and BPH32-NIL. BPH2 and BPH17-ptb showed resistance to antibiosis and antixenosis, while BPH17-ptb and BPH32 showed tolerance. These results contribute to the development of durable BPH resistance lines using three resistance genes from ‘PTB33’.     

Breeding cowpea for resistance to Striga gesnerioides in the Nigerian dry savannas using marker‐assisted selection

Historically, conventional breeding has been the primary strategy used to develop a number of Striga‐resistant varieties currently grown in the Sahel of Western Africa. In this study, we have successfully developed and applied a marker‐assisted selection strategy that employs a single backcross programme to introgress Striga resistance into farmer preferred varieties of cowpea for the Nigeria savannas. In this strategy, we have introduced the Striga resistance gene from the donor parent IT97K‐499‐35 into an elite farmer preferred cowpea cultivar ‘Borno Brown’. The selected 47 BC₁F₂ populations confirmed the recombinants with desirable progeny having Striga resistance gene(s). The 28 lines selected in the BC₁F_2:4 generation with large seed size, brown seed coat colour and carrying marker alleles were evaluated in the field for resistance to Striga resistance. This led to the selection of a number of desirable improved lines that were immune to Striga having local genetic background with higher yield than those of their parents and standard varieties.     

Mapping of quantitative trait loci associated with rice black‐streaked dwarf virus disease and its insect vector in rice (Oryza sativa L.)

Rice black‐streaked dwarf virus disease (RBSDVD), transmitted by small brown planthopper (SBPH, Laodelphax striatellus), causes serious loss in rice production. Breeding resistant cultivars are one of the most effective strategies to control the virus disease and its vector. By both natural inoculations in the field and modified seedling‐box screening test in the glasshouse, an indica variety WR24 showed high resistance to RBSDVD and SBPH. An F_2:3 population consisting of 153 lines derived from a cross between WR24 and a susceptible japonica variety Suyunuo was used for quantitative trait loci (QTL) analysis of RBSDVD and SBPH resistance. The linkage map consisting of 130 SSR markers was constructed with an average marker interval of 13.90 cM, spanning a total of 1890.9 cM. Totally, five QTLs for RBSDV resistance, viz. qRBSDV3^WR24, qRBSDV6^WR24, qRBSDV7^WR24, qRBSDV9^WR24 and qRBSDV11^WR24, were detected on chromosomes 3, 6, 7, 9 and 11, with LOD scores of 2.7, 3.08, 3.13, 5.28 and 3.7, respectively. Meanwhile, three QTLs for SBPH resistance, including qSBPH5^WR24, qSBPH7^WR24 and qSBPH10^WR24, were mapped on chromosomes 5, 7 and 10, with LOD scores of 2.18, 3.5 and 3.57, respectively. All resistant alleles were from WR24. Among these QTLs, qRBSDV7^WR24, qSBPH5^WR24 and qSBPH10^WR24 were newly reported, and qSBPH10^WR24 showed major effect that explained 17.9% of total phenotypic variance. The RBSDVD and SBPH resistance QTLs and the tightly linked DNA markers can be utilized in RBSDV and SBPH resistance breeding in rice.     

Breeding and characterization of the world’s first practical rice variety with resistance to brown spot (Bipolaris oryzae) bred using marker-assisted selection

Brown spot (BS) caused by Bipolaris oryzae is a serious disease of rice and decreases grain yield. Breeding for BS resistance is an economical solution but has not hitherto been achieved. To develop a practical BS-resistant variety, we introduced a chromosomal segment including a quantitative trait locus (QTL) for BS resistance, qBSfR11, derived from the BS-resistant local resource ‘Tadukan’, into the genetic background of the high-yielding but susceptible ‘Mienoyume’. Resistance is controlled by a single recessive gene in a 1.3-Mbp region. We named this gene bsr1 (brown spot resistance 1). The near-isogenic line bsr1-NIL had a greater yield with larger grain width than Mienoyume but similar other agronomic traits in fields where BS was mild; it had a significantly lower BS disease score and a 28.8% higher yield in fields where BS was more severe, and it showed resistance to multiple isolates of BS fungus. It showed stable resistance to BS and had excellent agricultural traits in the presence of BS. We developed the bsr1-NIL with resistance to BS and applied it for variety registration to Ministry of Agriculture, Forestry and Fisheries in Japan as ‘Mienoyume BSL’. This is the first report for the BS resistant rice variety bred using marker-assisted selection.     

A dominant gene in rice for resistance to white-backed planthopper and its relationship to other plant characteristics

Summary The inheritance of resistance to white-backed planthopper, Sogatella furcifera Horvath, was investigated in a rice, Oryza sativa L., cultivar N22. Resistance to the white-backed planthopper in the cross IR30×N22 appears to be governed by a single dominant gene-designated Wbph. The classification for various characteristics of 397 F₃ families of the IR30×N22 cross confirmed earlier results about the monogenic dominant control of resistance to brown planthopper, green leafhopper, and bacterial leaf blight, and about the monogenic recessive control of short stature. Additionally, the genes governing plant height and resistance to white-backed planthopper, brown planthopper, green leafhopper, and bacterial leaf blight were found to segregate independently of each other in these 397 F₃ families.     

Marker‐assisted breeding of Pi‐1 and Piz‐5 genes imparting resistance to rice blast in PRR78, restorer line of Pusa RH‐10 Basmati rice hybrid

Rice blast, caused by fungus Magnaporthe grisea, is a serious disease causing considerable economic damage worldwide. Best way to overcome disease is to breed for disease‐resistant cultivars/parental lines of hybrids. Pusa RH10, first aromatic, fine‐grain rice hybrid released and cultivated extensively in India. Hybrid and its parental lines, Pusa 6A and PRR78, are highly susceptible to blast. CO39 pyramid carrying two dominant, broad‐spectrum blast‐resistance genes, viz. Pi‐1 and Piz‐5, used as a donor parent to introgress these genes into PRR78 using marker‐assisted backcrossing (MABC). Microsatellite markers RM5926 and AP5659‐5 tightly linked to Pi‐1 and Piz‐5 genes, respectively, were used for foreground selection to derive introgression lines. Further, these lines were evaluated for agronomic performance, disease reaction and cooking quality traits along with PRR78. Most of the improved lines were on par with PRR78 for all traits evaluated except gelatinization temperature. Recurrent parent genome percentage (RPG) study also revealed similarity of these lines with PRR78. Hybrids derived using improved PRR78 lines were superior over Pusa RH10 in terms of yield.     

Marker‐assisted selection for rice blast resistance genes Pi2 and Pi9 through high‐resolution melting of a gene‐targeted amplicon

The use of host resistance (R) genes is considered the most cost‐effective option to control the rice blast disease. The two allelic R genes Pi2 and Pi9 confer very broad‐spectrum resistance against blast isolates collected worldwide. However, the two genes have not yet been widely deployed in rice breeding programmes. Availability of specific markers for them would facilitate incorporating the two R genes into new rice lines through marker‐assisted selection. Herein, we report the development and utilization of a robust and specific marker for the Pi2 and Pi9. This marker was derived from polymorphisms within the target gene, and achieved simultaneously distinguish Pi2 and Pi9 from other alleles through high‐resolution melting of a small amplicon. With the marker, we were able to transfer the Pi2 into an elite restorer line through marker‐assisted backcrossing, successfully obtained effective resistance to blast disease, and we were also able to, respectively, incorporate the Pi2 and Pi9 with two other R genes. As the additive effect, blast resistance in these stacking lines harbouring three R genes were significantly improved.