Showing papers in "Breeding Science in 2020"

Cassava breeding and agronomy in Asia: 50 years of history and future directions

Abstract: In Asia, cassava (Manihot esculenta) is cultivated by more than 8 million farmers, driving the rural economy of many countries. The International Center for Tropical Agriculture (CIAT), in partnership with national agricultural research institutes (NARIs), instigated breeding and agronomic research in Asia, 1983. The breeding program has successfully released high-yielding cultivars resulting in an average yield increase from 13.0 t ha-1 in 1996 to 21.3 t ha-1 in 2016, with significant economic benefits. Following the success in increasing yields, cassava breeding has turned its focus to higher-value traits, such as waxy cassava, to reach new market niches. More recently, building resistance to invasive pests and diseases has become a top priority due to the emergent threat of cassava mosaic disease (CMD). The agronomic research involves driving profitability with advanced technologies focusing on better agronomic management practices thereby maintaining sustainable production systems. Remote sensing technologies are being tested for trait discovery and large-scale field evaluation of cassava. In summary, cassava breeding in Asia is driven by a combination of food and market demand with technological innovations to increase the productivity. Further, exploration in the potential of data-driven agriculture is needed to empower researchers and producers for sustainable advancement.

Breeding buckwheat for nutritional quality.

Abstract: Common buckwheat (Fagopyrum esculentum Moench, CB) and Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn., TB) are used in human nutrition. The idea to screen in the haploid phase for genes affecting low amylose concentration opens the possibility for the effective search of low amylose (waxy) genotypes in CB populations. Self-pollinated homozygous plants of TB might allow us to use a part of endosperm for screening of amylose content. Phenolic substances have a significant inhibitory effect on the digestion of CB and TB proteins, thus metabolites may have impact on protein digestibility. Digestion-resistant peptides are largely responsible for the bile acid elimination. Breeding to diminish polyphenols and anti-nutritional substances might have negative effects on the resistance of plants against pests, diseases and UV-radiation. Bread and pasta are popular CB and TB dishes. During dough making most of CB or TB rutin is degraded to quercetin by rutin-degrading enzymes. The new trace-rutinosidase TB variety makes possible making TB bread with considerable amount of rutin, preserving the initial rutin from flour. Breeding CB and TB for larger embryos would make it possible to increase protein, rutin, and essential minerals concentration in CB and TB grain.

Present status and future perspectives of breeding for buckwheat quality.

Abstract: Buckwheat is an important crop globally. It has been processed into cereal grain, noodles, confectionery, bread, and fermented foods for many years. Buckwheat production and processing has supported local economies and is deeply related to the culture of some regions. Buckwheat has many unique traits as a food, with a good flavor and color. In addition, buckwheat is also a healthy food because it contains bioactive compounds that have anti-oxidative, anti-hypertensive, and anti-obesity properties. Therefore, breeding of buckwheat for quality is an important issue to be addressed. Compared to other crops, there is still a lack of basic information on quality, including bioactive compounds generation and enhancement. However, some mechanisms for modifying and improving the quality of buckwheat varieties have recently been identified. Further, some varieties with improved quality have recently been developed. In this review, we summarize the issues around buckwheat quality and review the present status and future potential of buckwheat breeding for quality.

Biosynthesis and regulation of flavonoids in buckwheat.

Abstract: Buckwheat contains an abundance of antioxidants such as polyphenols and is considered a functional food. Among polyphenols, flavonoids have multiple functions in various aspects of plant growth and in flower and leaf colors. Flavonoids have antioxidant properties, and are thought to prevent cancer and cardiovascular disease. Here, we summarize the flavonoids present in various organs and their synthesis in buckwheat. We discuss the use of this information to breed highly functional and high value cultivars.

Genomics-assisted breeding in minor and pseudo-cereals

Abstract: Minor and pseudo-cereals, which can grow with lower input and often produce specific nutrients compared to major cereal crops, are attracting worldwide attention. Since these crops generally have a large genetic diversity in a breeding population, rapid genetic improvement can be possible by the application of genomics-assisted breeding methods. In this review, we discuss studies related to biparental quantitative trait locus (QTL) mapping, genome-wide association study, and genomic selection for minor and pseudo-cereals. Especially, we focus on the current progress in a pseudo-cereal, buckwheat. Prospects for the practical utilization of genomics-assisted breeding in minor and pseudo-cereals are discussed including the issues to overcome especially for these crops.

Marker-assisted selection of qMrdd8 to improve maize resistance to rough dwarf disease.

Abstract: Maize rough dwarf disease (MRDD) is caused by viruses in the Fijivirus genus in the family Reoviridae. MRDD resistance can be improved by a combination of marker-assisted selection (MAS) and conventional breeding strategies. In our previous study, we fine-mapped a major QTL qMrdd8 and developed the functional Indel marker IDP25K. In the present study, qMrdd8 from the donor parent X178 was introgressed into elite inbred lines derived from the three corn heterotic groups using multi-generation backcrossing and MAS. Recipient lines included Huangzao4, Chang7-2, Ye478, Zheng58, Zhonghuang68, B73, and Ji846. Markers used for foreground selection included IDRQ4, IDRQ47, IDP25K, and IDP27K. Background selection was carried out in the BC3 or BC4 using 107 SSR markers to select lines with the highest rate of recovery of the particular recurrent parent genome. Plants from BC4F2 and BC3F2 that carried the shortest qMrdd8 interval from X178 and those with the highest rate of recovery of the recurrent parent genome were then selected to create converted homozygous inbred lines. In 2017, seven converted inbred lines and five hybrids exhibited enhanced resistance to MRDD, while other agronomic traits were not affected under nonpathogenic stress conditions. Thus, the MRDD resistance allele at the qMrdd8 locus, or IDP25K, should be valuable for maize breeding programs in China.

Important agronomic characteristics of yielding ability in common buckwheat; ecotype and ecological differentiation, preharvest sprouting resistance, shattering resistance, and lodging resistance

Abstract: Common buckwheat is recognized as a healthy food because its seed contains large amounts of protein, minerals, and rutin. However, the yielding ability of common buckwheat is lower than that of other major crops. The short growing period, moisture injury, occurrence of sterile seeds due to lack of flower-visiting insects, and yield loss due to lodging and shattering cause low and unstable grain yield. Therefore, many common buckwheat breeders have tried to increase yielding ability by improving various characteristics. Recently, new breeding objectives for improving yielding ability by increasing preharvest sprouting resistance; reducing shattering loss; introducing self-compatibility; the ecotype, and semidwarf have been reported. In this review, we introduce the research on the important agronomic characteristics, preharvest sprouting resistance, ecotype and ecological differentiation, shattering resistance, and lodging resistance in common buckwheat.

Classification and systematics of the Fagopyrum species.

Abstract: Fagopyrum (Polygonaceae) is a small genus including less than 30 species, mostly endemic to southern China. The genus includes two cultivated species, common buckwheat F. esculentum and Tartary buckwheat F. tataricum. Fagopyrum is clearly distinguished from other genus in Polygonaceae by the central position of embryo in achene. The genus is divided into two major groups, namely the cymosum group and the urophyllum group, based on morphological characters and molecular systematics. In the last three decades the number of species in the genus has doubled by the discovery of new species by Japanese and Chinese groups. Most of them are revealed to be included in the urophyllum group based on morphological and molecular genetic analyses. Molecular systematic surveys have also detected inappropriate treatment of some non-Fagopyrum species as new species or combination in the genus.

Identification of QTLs involved in cold tolerance during the germination and bud stages of rice (Oryza sativa L.) via a high-density genetic map

Abstract: Low-temperature tolerance during the germination and bud stages is an important characteristic of direct-seeded rice (DSR). Recombinant inbred lines (RILs) derived from indica rice H335, which is highly tolerant to low temperature, and indica rice CHA-1, which is sensitive to low temperature, were used to identify quantitative trait loci (QTLs) associated with low-temperature tolerance during the germination and bud stages. a total of 11 QTLs were detected based on a high-density genetic map; among these, six QTLs explained 5.13-9.42% of the total phenotypic variation explained (PVE) during the germination stage, and five QTLs explained 4.17-6.42% of the total PVE during the bud stage. All QTLs were distributed on chromosome 9, and all favourable alleles originated from H335. The physical position of each QTL was determined, and 11 QTLs were combined into five genetic loci; three of these loci are involved during the germination stage (loci 1, 2, and 3), and three are involved during the bud stage (loci 3, 4, and 5). Loci 2, 4 and 5 were repeatedly detected in the wet season (WS) and dry season (DS). Notably, loci 3 was detected during both the germination and bud stages. These loci are good candidates for future studies of gene function and could serve as highly valuable genetic factors for improving cold tolerance during the germination and bud stages of rice.

Fine mapping of a major locus representing the lack of prickles in eggplant revealed the availability of a 0.5-kb insertion/deletion for marker-assisted selection

Abstract: As prickles cause labour inefficiency during cultivation and scratches on the skin of fruits during transportation, they are considered undesirable traits of eggplant (Solanum melongena L.). Because the molecular basis of prickle emergence has not been entirely revealed in plants, we mapped an eggplant semi-dominant Prickle (Pl) gene locus, which causes the absence of prickles, on chromosome 6 of a linkage map of the F2 population derived from crossing the no-prickly cultivar 'Togenashi-senryo-nigo' and the prickly line LS1934. By performing synteny mapping with tomato, the genomic region corresponding to the eggplant Pl locus was identified. Through bacterial artificial chromosome (BAC) screening, positive BAC clones and the contig sequence that harbour the Pl locus in the prickly eggplant genome were revealed. The BAC contig length was 133 kb, and it contained 16 predicted genes. Among them, a characteristic 0.5-kb insertion/deletion was detected. As the 0.5-kb insertion was commonly identified with the prickly phenotype worldwide, a primer pair that amplifies the insertion/deletion could be used for marker-assisted selection of the no-prickly phenotype. Such findings contribute to map-based-cloning of the Pl gene and the understanding of gene function, ultimately providing new insights into the regulatory molecular mechanisms underlying prickle emergence in plants.

Understanding buckwheat allergies for the management of allergic reactions in humans and animals.

Abstract: Buckwheat allergy is an immediate hypersensitivity reaction that includes anaphylaxis mediated by specific IgE antibodies. Several IgE-binding proteins in common buckwheat have been reported to be possible clinically relevant buckwheat allergens. Although common buckwheat is popularly consumed in Asia, buckwheat allergy is becoming a serious problem not only in Asia but also in Europe. In addition, common buckwheat has also been found to be a causative agent of allergic symptoms in animals. In recent years, in addition to conventional food allergy testing methods, the development of component-resolved diagnosis (CRD) has improved the diagnostic accuracy of food allergy. The identification of allergens is essential for the construction of CRD. In this review, we introduce the different types of buckwheat allergens and discuss how each buckwheat allergen contributes to the diagnosis of buckwheat allergy. We also present the analysis of buckwheat allergen that will help reduce the allergenicity of common buckwheat and reduce buckwheat allergen molecules. These findings may be beneficial in overcoming buckwheat allergies in humans and animals.

Effect of salt tolerance on biomass production in a large population of sorghum accessions.

Abstract: Salinity causes major reductions in cultivated land area, crop productivity, and crop quality, and salt-tolerant crops have been required to sustain agriculture in salinized areas. The annual C4 crop plant Sorghum bicolor (L.) Moench is salt tolerant, with large variation among accessions. Sorghum's salt tolerance is often evaluated during early growth, but such evaluations are weakly related to overall performance. Here, we evaluated salt tolerance of 415 sorghum accessions grown in saline soil (0, 50, 100, and 150 mM NaCl) for 3 months. Some accessions produced up to 400 g per plant of biomass and showed no growth inhibition at 50 mM NaCl. Our analysis indicated that the genetic factors that affected biomass production under 100 mM salt stress were more different from those without salt stress, comparing to the differences between those under 50 mM and 100 mM salt stress. A genome-wide association study for salt tolerance identified two single-nucleotide polymorphisms (SNPs) that were significantly associated with biomass production, only at 50 mM NaCl. Additionally, two SNPs were significantly associated with salt tolerance index as an indicator for growth response of each accession to salt stress. Our results offer candidate genetic resources and SNP markers for breeding salt-tolerant sorghum.

Buckwheat heteromorphic self-incompatibility: genetics, genomics and application to breeding

Abstract: Common buckwheat (Fagopyrum esculentum Moench 2n = 2x = 16) is an outcrossing crop with heteromorphic self-incompatibility due to its distylous flowers, called pin and thrum. In pin plants, a long style is combined with short stamens and small pollen grains; in thrum plants, a short style is combined with long stamens and large pollen grains. Both the intra-morph self-incompatibility and flower morphology are controlled by a single genetic locus named the S locus; thrum plants are heterozygous (Ss) and pin plants are homozygous recessive (ss) at this locus. Self-incompatibility is an obstacle for establishing pure lines and fixation of agronomically useful genes. Elucidation of the molecular mechanism of heterostylous self-incompatibility of common buckwheat has continued for a quarter of a century. Recent advances in genomic and transcriptomic analyses using next-generation sequencing have made it possible to determine the genomic region harboring the buckwheat S locus and to identify novel genes at this locus. In this review, we summarize the current knowledge on buckwheat heterostyly gained from conventional and molecular genetics and genomics. We also discuss the application of these studies to breeding of common buckwheat.

Current status and prospects of common buckwheat breeding in Japan.

Abstract: In this review, the current status and prospects of common buckwheat (Fagopyrum esculentum Moench.) breeding in Japan are summarized. The varieties that have been registered in Japan so far are introduced with details regarding their breeding source populations and breeding methods. Because the main breeding method used for common buckwheat is mass selection, the merits and demerits of this method are explored from the perspective of heritability. Although there are many breeding objectives in common buckwheat, high yield and yield stability are discussed here. Regarding the potential of common buckwheat breeding in the future, the prospects of effective exploitation of self-fertility and selection based on genomic information are examined.

A single QTL on chromosome 6DS derived from a winter wheat cultivar 'OW104' confers resistance to Wheat yellow mosaic virus.

Abstract: Wheat yellow mosaic (WYM) is a soilborne disease caused by Wheat yellow mosaic virus (WYMV). Symptoms include yellow mosaic coloring of leaves, stunting, and growth inhibition. Severe infection may result in yield loss. WYM is one of the most serious diseases affecting wheat production in East Asia. The most effective control is through breeding resistant cultivars. A winter wheat cultivar, 'OW104', shows little to no symptoms in heavily WYMV-infested fields in Hokkaido, Japan. Here we detected Qym4, a QTL accounting for 45%-57% of WYMV resistance, in the vicinity of the markers Xcfd49, Xbarc183, and Xgpw4357 on wheat chromosome arm 6DS. F3 progenies with 'OW104' allele at Qym4 showed significantly higher resistance than those with 'Hokushin' homozygote or heterozygote. We developed 'Hokushin' near-isogenic lines by backcrossing with 'Hokushin' as the recurrent parent and 'OW104' as the resistance donor. All the WYMV-resistant BC5F1/BC4F1 plants carried 'OW104' allele only at Xcfd49. Our results suggest that the introduction of Qym4 confers resistance to WYMV in winter wheat.

Mikan Genome Database (MiGD): integrated database of genome annotation, genomic diversity, and CAPS marker information for mandarin molecular breeding.

Abstract: Citrus species are some of the most valuable and widely consumed fruits globally. The genome sequences of representative citrus (e.g., Citrus clementina, C. sinensis, C. grandis) species have been released but the research base for mandarin molecular breeding is still poor. We assembled the genomes of Citrus unshiu and Poncirus trifoliata, two important species for citrus industry in Japan, using hybrid de novo assembly of Illumina and PacBio sequence data, and developed the Mikan Genome Database (MiGD). The assembled genome sizes of C. unshiu and P. trifoliata are 346 and 292 Mb, respectively, similar to those of citrus species in public databases; they are predicted to possess 41,489 and 34,333 protein-coding genes in their draft genome sequences, with 9,642 and 8,377 specific genes when compared to C. clementina, respectively. MiGD is an integrated database of genome annotation, genetic diversity, and Cleaved Amplified Polymorphic Sequence (CAPS) marker information, with these contents being mutually linked by genes. MiGD facilitates access to genome sequences of interest from previously reported linkage maps through CAPS markers and obtains polymorphism information through the multiple genome browser TASUKE. The genomic resources in MiGD (https://mikan.dna.affrc.go.jp) could provide valuable information for mandarin molecular breeding in Japan.

Efficient transient gene expression system using buckwheat hypocotyl protoplasts for large-scale experiments

Abstract: Buckwheat (Fagopyrum esculentum) is cultivated worldwide and its flour is used in a variety of food products. Although functional analyses of genes in buckwheat are highly desired, reliable methods to do it have yet to be developed. In this study we established a simple and efficient transient gene expression system using buckwheat protoplasts isolated from young hypocotyls using 96-well plates as a high-throughput platform. The transformation efficiency was comparable with that of similar systems, such as Arabidopsis mesophyll protoplasts. Stable results were obtained in a typical example of the experiment to examine transcription factor activity. This system shows potential for the large-scale analysis of gene function using protoplast isolated from fewer and younger plants than the conventional system and may provide novel information for efficient buckwheat breeding.

A novel genotype DATTO5 developed using the five genes exhibits the fastest heading date designed in rice.

Abstract: The optimization of heading date is a key aspect for maximizing grain productivity in cereal crops including rice. The combinations of genes for heading date, a quantitative trait, are a major driver in the wide adaptability of cultivated rice worldwide. Here, we identified a novel QTL, qDTH3 (quantitative trait locus for days-to-heading on chromosome 3), for early flowering time in the F2 population derived from a cross between Hoshinoyume (HS) and Daichinohoshi (DH) among local rice populations with extremely early heading date. The DH allele at qDTH3, qDTH3DH , headed 2.7 days earlier than the HS allele at qDTH3, qDTH3HS . We sought to design a genotype for earlier heading date by pyramiding of five heading date genes. We designated this aggregate of the five genes as DATTO5. Plants with DATTO5 were selected from the F2 population derived from a cross between DH and HShd5, which is a near-isogenic line carrying a loss-of-function of days to heading 8 in a genetic background of HS. Plants with DATTO5 exhibited earlier heading date but reduced fitness, including shorter culm and panicle length and fewer seeds compared with HS, as a representative local rice variety with extremely early heading date.

Days to heading, controlled by the heading date genes, Hd1 and DTH8, limits rice yield-related traits in Hokkaido, Japan

Abstract: A key aspect of rice breeding programs is the optimization of days to heading (DTH) for maximizing grain productivity in cultivation areas. Here, the effects of genotypes for heading date on yield-related traits in rice (culm and panicle length (CL and PL), panicle number (PN), and total number of seeds) were investigated. Heading date 1 (Hd1) and Days to heading 8 (DTH8) are the main controllers of the variation in heading date in the rice population of Hokkaido, Japan. Thus, an F2 population (n = 192) derived from a cross between Kitaibuki (Hd1dth8) and Akage (hd1DTH8) was developed. Significant differences in DTH were found among all combinations. Each genotype for heading date showed variations in the yield-related traits without a significant difference. However, DTH exhibited high positive coefficient values (more than 0.709) with the yield-related traits except for PN, which had a negative coefficient value of -0.431. A later heading date resulted in a longer growth duration and a higher yield with a combination of longer PL and CL and lower PN. These results suggest that DTH limits the yield-related traits rather than the genotype for heading date.

gw2 mutation increases grain width and culm thickness in rice (Oryza sativa L.).

Abstract: Grain size is one of the most important agricultural traits in rice. To increase grain yield, we screened a large grain mutant from mutants with the 'Koshihikari' background. As a result, we obtained a mutant, KEMS39, that has a large grain size and increased yield. Cultivation tests revealed that this mutant had improved lodging resistance with thicker internodes. Next-generation sequencing analysis revealed the presence of a 67 bp deletion in the GW2 mRNA, owing to a mutation in the 3' splice site of the sixth intron of the GW2 gene. To determine whether this mutation was responsible for the larger grain and thicker internodes, we performed gene editing and obtained a mutant with a 7 bp deletion, including this 3' splice site. As this gw2 mutant had large grains and thicker internodes, the causal gene of KEMS39 was determined as GW2. Thicker internodes are attributed to the pleiotropic effect of gw2 mutation. On the basis of these results, we conclude that gw2 mutation has the potential to be an important genetic resource with the ability to achieve a well-balanced and high-yielding effect that simultaneously improves grain productivity and lodging resistance.

Low-cost RNA extraction method for highly scalable transcriptome studies.

Abstract: RNA extraction has been improved by integration of a variety of materials in the protocol, such as phenol, guanidine thiocyanate, and silica, according to the case-specific demands. However, few methods have been designed for high-throughput RNA preparation for large-scale transcriptome studies. In this study, we established a high-throughput guanidinium thiocyanate and isopropyl alcohol based RNA extraction method (HighGI). HighGI is based on simple and phenol-free homemade buffers and the cost is substantially lower than a column-based commercial kit. We demonstrated that the quality and quantity of RNA extracted with HighGI were comparable to those extracted with a conventional phenol/chloroform-based method and a column-based commercial kit. HighGI retained small RNAs less than 200 bp, which are lost with a commercial column-based kit. We also demonstrated that HighGI is readily applicable to semi-automated RNA extraction. HighGI enables high-throughput RNA extraction for large-scale RNA preparation with high yield and quality.

Development of co-dominant markers linked to a hemizygous region that is related to the self-compatibility locus (S) in buckwheat (Fagopyrum esculentum).

Abstract: Common buckwheat (Fagopyrum esculentum) is a heterostylous self-incompatible (SI) species with two different flower morphologies, pin and thrum. The SI trait is controlled by a single gene complex locus, S. Self-compatible (SC) lines were developed by crossing F. esculentum and F. homotropicum; these lines have an SC gene, Sh , which is dominant over the s allele and recessive to the S allele. S-ELF3 has been identified as a candidate gene in the S locus and is present in the S and Sh but not s alleles. A single-nucleotide deletion in the S-ELF3 gene of the Sh allele results in a frame shift. To develop co-dominant markers to distinguish between ShSh and Shs plants, we performed a next-generation sequencing analysis in combination with bulked-segregant analysis. We developed four co-dominant markers linked to the S locus. We investigated the polymorphism frequency between a self-compatible line and leading Japanese buckwheat cultivars. Linkage between a developed sequence-tagged-site marker and flower morphology was confirmed using more than 1000 segregating plants and showed no recombination. The developed markers would be useful for buckwheat breeding and also to produce lines for genetic analysis such as recombinant inbred lines.

ISSR markers to assess genetic diversity of cultivated populations from artificial selection of Stevia rebaudiana (Bert.) Bertoni.

Abstract: Artificial selection related with important agronomic characteristics of Stevia rebaudiana may cause genetic divergence and formation of genetically structured populations with genetic uniformity or diversity within cultivars. Current study employed inter simple sequence repeats of DNA (ISSR markers) to assess genetic diversity within and among a single cultivated population maintained through sexual propagation (SR1) and four cultivated populations generated by artificial selection and maintained by vegetative propagation (SR2-SR5). Highest polymorphism rate was reported in SR1 (89.24%), whilst the lowest rate of polymorphism occurred in SR2 (60.13%). ISSR markers revealed that selection of plants with traits of vegetative-propagated interest may lead towards the generation of genetically more uniform DNA-level populations, while plants maintained by sexual propagation have high genetic variability. High estimated genetic divergence level indicated that the five areas of stevia form genetically structured populations. SR2 and SR4 are constituted by plants more homogeneous at DNA level for the selected characteristics than plants of SR3 and SR5 populations. Predominant and homogeneous genotypes selected at SR2 and SR4 areas could be valuable for tracing strategies to obtain stevia plants with the desirable agronomic characteristics through crosses between contrasting individuals in future breeding programs.

Assessment of genetic diversity in Camellia oleifera Abel. accessions using morphological traits and simple sequence repeat (SSR) markers.

Abstract: Camellia oleifera Abel. (C. oleifera) is a cultivable plant with important economic value. It is very helpful for the scientific utilization, cultivation and preservation of germplasm resources through evaluating the genetic diversity. In this study, we estimated the genetic relationship of 150 accessions of C. oleifera using morphological and economic traits, as well as SSR molecular marker. Through the variation and cluster analysis of 17 morphological and economic traits, the germplasm was divided into a candidate core breeding group with higher economic traits and a core breeding group with higher morphological traits. The genetic similarity coefficients of SSR markers ranged from 0.05 to 0.91, and the germplasm materials were divided into five groups. The results demonstrated that C. oleifera germplasms perform a rich genetic variation. This is the first report to evaluate the genetic diversity of different C. oleifera germplasms using the morphological and economic traits, together with SSR molecular marker, and the results allow us to find evidence for the origin of varieties, establish core breeding populations and its fingerprint.

History of the progressive development of genetic marker systems for common buckwheat.

Abstract: Genotyping is an essential procedure for identifying agronomically useful genes and analyzing population structure. Various types of genetic marker systems have been developed in common buckwheat (Fagopyrum esculentum Moench). In the 1980s, morphological and allozyme markers were used to construct linkage maps. Until the early 2000s, allozyme markers were widely used in population genetics studies. Such studies demonstrated that cultivated common buckwheat likely originated in the Sanjiang area of China. In the late 1990s and early 2000s, advances in PCR technology led to the development of various DNA marker systems for use in linkage mapping. However, PCR-based markers did not completely cover the genome, making genetic analysis of buckwheat challenging. The subsequent development of next generation sequencing, a game-changing technology, has allowed genome-wide analysis to be performed for many species. Indeed, 8,884 markers spanning 756 loci were recently mapped onto eight linkage groups of common buckwheat; these markers were successfully used for genomic selection to increase yield. Furthermore, draft genome sequences are now available in the Buckwheat Genome DataBase (BGDB). In this review, I summarize advances in the breeding and genetic analysis of common buckwheat based on contemporary genetic marker systems.

TaqMan-MGB SNP genotyping assay to identify 48 citrus cultivars distributed in the Japanese market.

Abstract: A citrus cultivar identification system using CAPS marker has been developed on nursery trees, but this needs to be extended to include various product types, such as imported fruits and processed products. Here, we developed a new cultivar identification system using TaqMan-MGB SNP genotyping assay. Eight probe and primer sets were designed to amplify PCR fragments <100 bp to enable the genotyping of fresh and processed fruits in which predicted that insufficient quantities of DNA and residual impurities in the DNA extracts. The TaqMan-MGB SNP genotyping assay was stable and reproducible, and were confirmed to apply various sample sources, including leaves, fresh fruit, juice, canned fruit, and dry fruit. They could provide at least a single differentiating SNP to discriminate any paired combination among 48 citrus cultivars. Minimal marker subsets to identify the target cultivar were listed for each of 18 registered cultivars with valid patent. The allelic SNP genotypes of 48 citrus cultivars, which cover more than 98% of all citrus fruit shipment produced in Japan, is valuable for the referencing information in the DNA-based identification for fresh and processed fruits. This identification system will help protect registered cultivars and facilitate food fraud inspections.

Current status of genomic resources on wild relatives of rice

Abstract: Rice is a food crop of global importance, cultivated in diverse agro-climatic zones of the world. However, in the process of domestication many beneficial alleles have been eroded from the gene pool of the rice cultivated globally and eventually has made it vulnerable to a plethora of stresses. In contrast, the wild relatives of rice, despite being agronomically inferior, have inherited a potential of surviving in a range of geographical habitats. These adaptations enrich them with novel traits that upon introgression to modern cultivated varieties offer tremendous potential of increasing yield and adaptability. But, due to the unavailability of their genetic as well as genomic resources, identification and characterisation of these novel beneficial alleles has been a challenging task. Nevertheless, with the unprecedented surge in the area of conservation genomics, researchers have now shifted their focus towards these natural repositories of beneficial traits. Presently, there are several generic and specialized databases harboring genome-wide information on wild species of rice, and are acting as a useful resource for identification of novel genes and alleles, designing of molecular markers, comparative analysis and evolutionary biology studies. In this review, we introduce the key features of these databases focusing on their utility in rice breeding programs.

Development of diagnostic molecular markers for marker-assisted breeding against bacterial wilt in tomato.

Abstract: Bacterial wilt, caused by the Ralstonia pseudosolanacearum species complex, is an important vascular disease that limits tomato production in tropical and subtropical regions. Two major quantitative trait loci (QTL) of bacterial wilt resistance on chromosome 6 (Bwr-6) and 12 (Bwr-12) were previously identified in Solanum lycopersicum 'Hawaii 7996'; however, marker-assisted breeding for bacterial wilt resistance is not well established. To dissect the QTL, six cleaved amplified polymorphic sites (CAPS) and derived CAPS (dCAPS) markers within the Bwr-6 region and one dCAPS marker near Bwr-12 were developed, and resistance levels in 117 tomato cultivars were evaluated. Two markers, RsR6-5 on chromosome 6 and RsR12-1 on chromosome 12, were selected based on the genotypic and phenotypic analysis. The combination of RsR6-5 and RsR12-1 effectively distinguishes resistant and susceptible cultivars. Furthermore, the efficiency of the two markers was validated in the F3 generation derived from the F2 population between E6203 (susceptible) and Hawaii 7998 (resistant). Resistant alleles at both loci led to the resistance to bacterial wilt. These markers will facilitate marker-assisted breeding of tomato resistant to bacterial wilt.

Reduced susceptibility to a tobacco bushy top virus malawi isolate by loss of function in host eif(Iso)4e genes

Abstract: Tobacco bushy top disease (TBTD) is a viral disease of tobacco (Nicotiana tabacum L.) caused by mixed infection of Tobacco bushy top virus or Ethiopian tobacco bushy top virus and a helper virus. Despite its damage to tobacco, practical genetic resources for disease resistance have not been found. Here, we report that a mutation of tobacco eIF(iso)4E genes (eIF(iso)4E-S and eIF(iso)4E-T), which encode eukaryotic translation initiation factors, confers resistance (reduced susceptibility) to TBTD caused by a virus from Malawi (designated as tobacco bushy top virus Malawi isolate, TBTV-MW). RNAi lines in which eIF(iso)4E genes were silenced showed reduced susceptibility to TBTV-MW. We also tested chemically-induced single (eIF(iso)4E-S or eIF(iso)4E-T) and double (eIF(iso)4E-S and eIF(iso)4E-T) nonsense mutants for resistance to TBTV-MW. Suppression of eIF(iso)4E-S showed reduced susceptibility, and the resistance of the double mutant tended to be even stronger. eIF(iso)4E mutants also showed reduced susceptibility to TBTV-MW transmitted by aphids. To the best of our knowledge, the eIF(iso)4E-S mutant is the first genetic resource for TBTD resistance breeding in tobacco.

Breeding for three-line japonica hybrid rice combinations with high resistant starch content using molecular marker-assisted selection.

Abstract: Resistant starch (RS) is beneficial for human health, and especially for diabetics. Considering the high cost and low productivity of the Jiangtangdao 1 rice variety with high RS content, breeding high RS rice varieties exhibiting high productivity is essential. A molecular marker-assisted selection strategy was applied to increase RS content in a three-line hybrid rice variety. The functional rice variety Jiangtangdao 1, which contains sbe3-rs (on chr2) that controls the RS content, was used as the high RS content donor parent. Subsequently, male sterile maintainer and restorer lines containing homozygous sbe3-rs were bred using molecular marker-assisted selection combined with traditional breeding methods. The male sterile line was crossed with the restorer lines to identify the optimal hybrid combination with a high RS content. We obtained four combinations for which the yields were >50% higher than those of the control Jiangtangdao 1. In addition, there was no significant difference in the RS content between the combinations and Jiangtangdao 1. The hybrid rice plants with high RS content exhibited favorable agronomic traits and therefore have broad prospects for commercial application.