Gene Pyramiding Research Articles

Pyramiding ascorbate-glutathione pathway in Lycopersicum esculentum confers tolerance to drought and salinity stress.

Stacking Glutathione-Ascorbate pathway genes (PgSOD, PgAPX, PgGR, PgDHAR and PgMDHAR) under stress inducible promoter RD29A imparts significant tolerance to drought and salinity stress in Solanum lycopersicum. Although the exposure of plants to different environmental stresses results in overproduction of reactive oxygen species (ROS), many plants have developed some unique systems to alleviate the ROS production and mitigate its deleterious effect. One of the key pathways that gets activated in plants is ascorbate glutathione (AsA-GSH) pathway. To demonstrate the effect of this pathway in tomato, we developed the AsA-GSH overexpression lines by stacking the genes of the AsA-GSH pathway genes isolated from Pennisetum glaucoma (Pg) including PgSOD, PgAPX, PgGR, PgDHAR and PgMDHAR under stress inducible promoter RD29A. The overexpression lines have an improved germination and seedling growth with concomitant elevation in the survival rate. The exposure of transgenic seedlings to varying stress regiments exhibited escalation in the antioxidant enzyme activity and lesser membrane damage as reflected by decreased electrolytic leakage and little accumulation of malondialdehyde and H2O2. Furthermore, the transgenic lines accumulated high levels of osmoprotectants with increase in the relative water content. The increased photosynthetic activity and enhanced gaseous exchange parameters further confirmed the enhanced tolerance of AsA-GSH overexpression lines. We concluded that pyramiding of AsA-GSH pathway genes is an effective strategy for developing stress resistant crops.

Identification of Root Rot Resistance QTLs in Pea Using Fusarium solani f. sp. pisi-Responsive Differentially Expressed Genes.

Pisum sativum (pea) yields in the United States have declined significantly over the last decades, predominantly due to susceptibility to root rot diseases. One of the main causal agents of root rot is the fungus Fusarium solani f. sp. pisi (Fsp), leading to yield losses ranging from 15 to 60%. Determining and subsequently incorporating the genetic basis for resistance in new cultivars offers one of the best solutions to control this pathogen; however, no green-seeded pea cultivars with complete resistance to Fsp have been identified. To date, only partial levels of resistance to Fsp has been identified among pea genotypes. SNPs mined from Fsp-responsive differentially expressed genes (DEGs) identified in a preceding study were utilized to identify QTLs associated with Fsp resistance using composite interval mapping in two recombinant inbred line (RIL) populations segregating for partial root rot resistance. A total of 769 DEGs with single nucleotide polymorphisms (SNPs) were identified, and the putative SNPs were evaluated for being polymorphic across four partially resistant and four susceptible P. sativum genotypes. The SNPs with validated polymorphisms were used to screen two RIL populations using two phenotypic criteria: root disease severity and plant height. One QTL, WB.Fsp-Ps 5.1 that mapped to chromosome 5 explained 14.8% of the variance with a confidence interval of 10.4 cM. The other four QTLs located on chromosomes 2, 3, and 5, explained 5.3–8.1% of the variance. The use of SNPs derived from Fsp-responsive DEGs for QTL mapping proved to be an efficient way to identify molecular markers associated with Fsp resistance in pea. These QTLs are potential candidates for marker-assisted selection and gene pyramiding to obtain high levels of partial resistance in pea cultivars to combat root rot caused by Fsp.

Pyramiding of scald resistance genes in four spring barley MAGIC populations

Genome-Wide Association Studies (GWAS) of four Multi-parent Advanced Generation Inter-Cross (MAGIC) populations identified nine regions on chromosomes 1H, 3H, 4H, 5H, 6H and 7H associated with resistance against barley scald disease. Three of these regions are putatively novel resistance Quantitative Trait Loci (QTL). Barley scald is caused by Rhynchosporium commune, one of the most important barley leaf diseases that are prevalent in most barley-growing regions. Up to 40% yield losses can occur in susceptible barley cultivars. Four MAGIC populations were generated in a Nordic Public–Private Pre-breeding of spring barley project (PPP Barley) to introduce resistance to several important diseases. Here, these MAGIC populations consisting of six to eight founders each were tested for scald resistance in field trials in Finland and Iceland. Eight different model covariate combinations were compared for GWAS studies, and the models that deviated the least from the expected p-values were selected. For all QTL, candidate genes were identified that are predicted to be involved in pathogen defence. The MAGIC progenies contained new haplotypes of significant SNP-markers with high resistance levels. The lines with successfully pyramided resistance against scald and mildew and the significant markers are now distributed among Nordic plant breeders and will benefit development of disease-resistant cultivars.

The Impact of Climate Change on the Resistance of Rice Near-Isogenic Lines with Resistance Genes Against Brown Planthopper

BackgroundThe impact of climate change on insect resistance genes is elusive. Hence, we investigated the responses of rice near-isogenic lines (NILs) that carry resistance genes against brown planthopper (BPH) under different environmental conditions.ResultsWe tested these NILs under three environmental settings (the atmospheric temperature with corresponding carbon dioxide at the ambient, year 2050 and year 2100) based on the Intergovernmental Panel on Climate Change prediction. Comparing between different environments, two of nine NILs that carried a single BPH-resistant gene maintained their resistance under the environmental changes, whereas two of three NILs showed gene pyramiding with two maintained BPH resistance genes despite the environmental changes. In addition, two NILs (NIL-BPH17 and NIL-BPH20) were examined in their antibiosis and antixenosis effects under these environmental changes. BPH showed different responses to these two NILs, where the inhibitory effect of NIL-BPH17 on the BPH growth and development was unaffected, while NIL-BPH20 may have lost its resistance during the environmental changes.ConclusionOur results indicate that BPH resistance genes could be affected by climate change. NIL-BPH17 has a strong inhibitory effect on BPH feeding on phloem and would be unaffected by environmental changes, while NIL-BPH20 would lose its ability during the environmental changes.

Maize streak virus research in Africa: an end or a crossroad.

The economic importance of the maize streak virus disease to the African maize production dynamic is to be appreciated now more than ever due to the preponderant influence of a changing climate. Continued dependence on a single major-effect quantitative trait locus (QTL) called Msv1 on Chromosome 1 of Maize (Zea mays L.) is not guaranteed to ensure durable resistance to the causal pathogen. With over ten decades of research on the disease and its associated host plant resistance mechanisms, it is pertinent to consider future approaches to attaining durability by looking to the synergistic roles of moderate- and minor-effect QTLs located on other chromosomes so as to facilitate a secure farming system for sub-Saharan Africa. For this review, more than 40 publications relating to maize streak disease research were methodically analysed with about 30% making specific reference to conventional, molecular and transgenic approaches employed in introgressing, maintaining and improving streak resistance in maize. A meta-analysis of mapped QTLs conferring streak resistance was conducted in a bid to reveal any inter-dependence or co-localization of resistant loci and to aid decision-making for marker-assisted breeding. With the changing climatic conditions around the globe, man's preparedness in the event of an epidemic following any evolutionary process in the streak viral genome was determined as insufficient. Modern breeding approaches including gene pyramiding that could be considered in maize breeding programmes to ensure durability for streak resistance were proposed while improving maize for other abiotic stress tolerance, particularly drought.

Genetic interrelationships of elite sorghum (Sorghum bicolor (L.) Moench) genotypes bred for Striga resistance and compatibility with Fusarium oxysporum f. sp. strigae (FOS) revealed by microsatellites

ABSTRACT Genetic diversity is key for breeding population development, hybrid breeding, heterotic grouping and genetic analysis, among others. The objective of this study was to examine genetic interrelationships among elite sorghum genotypes bred for resistance to Striga hermonthica Del. Benth and S. asiatica (L.) Kuntze and compatible with a biological control agent Fusarium oxysporum f. sp. strigae (FOS) against Striga using 22 polymorphic simple sequence repeat (SSR) or microsatellite markers. The number of alleles per locus detected by the markers varied from 2 to 7 with a mean value of 4.23. The polymorphic information content (PIC) of the SSR markers ranged from 0.15 to 0.84 with a mean of 0.57 showing moderate discriminatory power of the primers. The neighbour-joining clustering algorithm classified the genotypes into three main groups, revealing narrow to moderate variations existing among the test genotypes. Overall, the genetic diversity revealed by the study will be useful for designed crosses and pyramiding of broad-based resistance genes to control Striga for sustainable sorghum production.

Molecular Breeding Strategy and Challenges Towards Improvement of Downy Mildew Resistance in Cauliflower (Brassica oleracea var. botrytis L.).

Cauliflower (Brassica oleracea var. botrytis L.) is one of the important, nutritious and healthy vegetable crops grown and consumed worldwide. But its production is constrained by several destructive fungal diseases and most importantly, downy mildew leading to severe yield and quality losses. For sustainable cauliflower production, developing resistant varieties/hybrids with durable resistance against broad-spectrum of pathogens is the best strategy for a long term and reliable solution. Identification of novel resistant resources, knowledge of the genetics of resistance, mapping and cloning of resistance QTLs and identification of candidate genes would facilitate molecular breeding for disease resistance in cauliflower. Advent of next-generation sequencing technologies (NGS) and publishing of draft genome sequence of cauliflower has opened the flood gate for new possibilities to develop enormous amount of genomic resources leading to mapping and cloning of resistance QTLs. In cauliflower, several molecular breeding approaches such as QTL mapping, marker-assisted backcrossing, gene pyramiding have been carried out to develop new resistant cultivars. Marker-assisted selection (MAS) would be beneficial in improving the precision in the selection of improved cultivars against multiple pathogens. This comprehensive review emphasizes the fascinating recent advances made in the application of molecular breeding approach for resistance against an important pathogen; Downy Mildew (Hyaloperonospora parasitica) affecting cauliflower and Brassica oleracea crops and highlights the QTLs identified imparting resistance against this pathogen. We have also emphasized the critical research areas as future perspectives to bridge the gap between availability of genomic resources and its utility in identifying resistance genes/QTLs to breed downy mildew resistant cultivars. Additionally, we have also discussed the challenges and the way forward to realize the full potential of molecular breeding for downy mildew resistance by integrating marker technology with conventional breeding in the post-genomics era. All this information will undoubtedly provide new insights to the researchers in formulating future breeding strategies in cauliflower to develop durable resistant cultivars against the major pathogens in general and downy mildew in particular.

Gene stacking: Approach of genetic engineering

Gene stacking is the process of addition of two or more gene of interest into a single plant. The combination or stacking of different traits or genes in plants is rapidly gaining popularity in biotech crop production. The new evolved trait is known as stacked trait and the crop is known as biotech stacked or simply stacked. This can be accomplished in many ways, one of which is gene pyramiding. Biotech stacks give crops a larger genetic and agronomic boost, allowing them to perform better in challenging farming situations. Biotech stacks are designed to increase productivity by overcoming biotic and abiotic challenges like as insect pests, diseases, weeds, and environmental stress. This review will explain about the gene stacking principle, the need for biotech stacking, and the many gene stacking methods.

New Simple Sequence Repeat Markers on Linkage Groups 2 and 7, and Investigation of New Sources of Eastern Filbert Blight Resistance in Hazelnut

Eastern filbert blight (EFB), caused by Anisogramma anomala, is a fungal disease threatening the european hazelnut (Corylus avellana) industry in the Willamette Valley of Oregon. The pathogen is endemic to the eastern United States where it causes little damage to the wild Corylus americana but causes severe cankers on most cultivars of the commercially important european hazelnut. The host genetic resistance in ‘Gasaway’ is conferred by a dominant allele at a single locus on linkage group 6 (LG6), and resistance from several other sources has been mapped to the same region. Some fungal isolates can overcome ‘Gasaway’ resistance, prompting a search for other sources of resistance. Resistance from other sources has been mapped to LG2 and LG7, for which additional simple sequence repeat (SSR) markers would facilitate marker-assisted selection (MAS). In this study, an in silico approach was used to develop new polymorphic SSR markers in the EFB resistance regions on LG2 and LG7. Starting with a search of 17 contigs of the ‘Jefferson’ genome sequence, 45 new polymorphic SSR markers were developed, characterized, and placed on the linkage map. The new SSR markers had an average of 10.18 alleles per locus, and average values for expected heterozygosity, observed heterozygosity, polymorphism information content, and frequency of null alleles of 0.72, 0.65, 0.68, and 0.068, respectively. Of the 42 new polymorphic SSRs segregating in the mapping population, 24 were on LG2, 12 were on LG7, and six were placed on other LGs. The new and previously developed SSR markers were used to study six new sources of EFB resistance, four from Russia and two from Crimea. Six resistant selections were crossed with susceptible selections, resulting in 7 progenies. Phenotyping for disease response revealed that segregation in progenies of the two Moscow selections (#2 and #27), one Russian selection (OSU 1187.101), and one Crimean selection (H3R12P62) fit the 1:1 segregation ratio expected for control of resistance by a dominant allele at a single locus; but in progenies of the other Russian selection (OSU 1166.123) and the other Crimean selection (H3R07P11), there was an excess of resistant seedlings. Correlation of disease scores and alleles at SSR loci indicated that resistance from three Russian selections (Moscow selections #2 and #27 and OSU 1166.123) and the Crimean selection H3R12P62 was on LG7, while resistance from Russian selection OSU 1187.101 was on LG2. Resistance from Crimean selection H3R07P11 was not correlated with markers on LG6, or LG2, or LG7. These sources and new SSR markers will be useful in MAS and the pyramiding of resistance genes in the breeding of new EFB-resistant cultivars.

Insights Into Natural Genetic Resistance to Rice Yellow Mottle Virus and Implications on Breeding for Durable Resistance.

Rice is the main food crop for people in low- and lower-middle-income countries in Asia and sub-Saharan Africa (SSA). Since 1982, there has been a significant increase in the demand for rice in SSA, and its growing importance is reflected in the national strategic food security plans of several countries in the region. However, several abiotic and biotic factors undermine efforts to meet this demand. Rice yellow mottle virus (RYMV) caused by Solemoviridae is a major biotic factor affecting rice production and continues to be an important pathogen in SSA. To date, six pathogenic strains have been reported. RYMV infects rice plants through wounds and rice feeding vectors. Once inside the plant cells, viral genome-linked protein is required to bind to the rice translation initiation factor [eIF(iso)4G1] for a compatible interaction. The development of resistant cultivars that can interrupt this interaction is the most effective method to manage this disease. Three resistance genes are recognized to limit RYMV virulence in rice, some of which have nonsynonymous single mutations or short deletions in the core domain of eIF(iso)4G1 that impair viral host interaction. However, deployment of these resistance genes using conventional methods has proved slow and tedious. Molecular approaches are expected to be an alternative to facilitate gene introgression and/or pyramiding and rapid deployment of these resistance genes into elite cultivars. In this review, we summarize the knowledge on molecular genetics of RYMV-rice interaction, with emphasis on host plant resistance. In addition, we provide strategies for sustainable utilization of the novel resistant sources. This knowledge is expected to guide breeding programs in the development and deployment of RYMV resistant rice varieties.

Leaf Rust: A Serious Disease of Wheat in Pakistan

Wheat is one of the major crops and also staple food of many areas. It fulfils the basic nutrition requirements of people. Demand of food in directly proportional to increasing number of populations. But most of the time yield is decreased due to attack of diseases. Wheat leaf rust is a common disease of wheat and cause 50% losses in yield. Leaf rust is cause by fungi named Puccinia triticina. Pathogen mostly attack on leaf blade and cause damage. Pathogen spores travel through wind and can spread on wide area. Many molecular studies have been for identification of resistant gene in cultivated and wild species to boost up the immunity of cultivated species against disease. For the purpose of genetic dissection scientist used molecular techniques like MAS and gene pyramiding, they used markers for identification of resistant genes. Scientists got success in their findings and identified many resistant gene in wild and cultivated varieties of wheat. Use of these genes will be helpful in breeding programs for the development of resistant varieties and to make increase in production.

Identification, pyramid and candidate genes of QTLs for associated traits based on a dense erect panicle rice CSSL-Z749 and five SSSLs, three DSSLs and one TSSL

BackgroundSeed-set density is an important agronomic trait in rice. However, its genetic mechanism is complex. Chromosome segment substitution lines (CSSLs) are ideal materials for studying complex traits.ResultsA rice CSSL, Z749, with a dense and erect panicle phenotype, was identified among progeny of the recipient parent Nipponbare and the donor parent Xihui 18. Z749 carried seven substitution segments (average length 2.12 Mb). Compared with Nipponbare, Z749 showed significant increases in the numbers of primary (NPB) and secondary branches (NSB), number of spikelets (SPP) and grains per panicle (GPP), seed-set density (SSD), and decrease in panicle length (PL). A secondary F2 population derived from a cross between Nipponbare and Z749 was used to map quantitative trait loci (QTLs) for associated traits. Fifteen QTLs distributed on chromosomes 5, 7, 8, and 10 were detected. The QTL qPL7 might be an allele of OsFAD8 and the remaining 14 QTLs (e.g., qSSD5 and qSSD10 etc.) might be novel. Fourteen QTLs were verified using five single-segment substitution lines (SSSLs). The seed-set density of Z749 was controlled predominantly by one major QTL (qSSD10) and two minor QTLs (qSSD5 and qSSD8). The QTLs qSSD10, qSSD5, and qSSD8 were fine-mapped to intervals of 1.05, 1.46, and 1.53 Mb on chromosomes 10, 5, and 8, respectively. Analysis of QTL additive effects indicated that qSSD5, qSSD8, and qSSD10 from Xihui18 increased seed-set density of Z749 by 14.10, 11.38, and 5.11 spikelets per 10 cm panicle, respectively. Analysis of QTL epistatic effects revealed that pyramiding of qSSD5 and qSSD8, qSSD5 and qSSD10, qSSD8 and qSSD10, and qSSD5, qSSD8 and qSSD10 produced novel genotypes with increased seed-set density.ConclusionsInheritance of seed-set density in Z749 was controlled predominantly by one major QTL (qSSD10) and two minor QTLs (qSSD5 and qSSD8). Then, they were fine-mapped to intervals of 1.05, 1.46, and 1.53 Mb on chromosomes 10, 5, 8, respectively. Two MAPK genes (OsMPK9 and OsMPK17) and one gene (candidate gene 6) involved in auxin metabolism might be candidate genes for qSSD5, and OsSAUR32 might be the candidate gene for qSSD8. Pyramiding of qSSD5, qSSD8, and qSSD10 enhanced seed-set density.

New Sources of Eastern Filbert Blight Resistance and Simple Sequence Repeat Markers on Linkage Group 6 in Hazelnut (Corylus avellana L.).

Commercial production of hazelnut (Corylus avellana) in Oregon’s Willamette Valley is threatened by eastern filbert blight (EFB), a serious canker disease caused by the pyrenomycete Anisogramma anomala (Peck) E. Müller. The fungus also prevents the establishment of hazelnut orchards in eastern North America. Genetic resistance is considered the most effective way to control the disease. A high level of EFB resistance was first discovered in ’Gasaway’. This resistance is conferred by a dominant allele at a single locus on linkage group 6 (LG6). Resistance from several additional sources has been assigned to the same chromosomal region. In this study, new simple sequence repeat (SSR) markers were developed for the resistance region on LG6 and new sources of resistance were investigated. Forty-two new SSR markers were developed from four contigs in the genome sequence of ‘Jefferson’ hazelnut, characterized, and nine of them were placed on LG6 of the genetic map. Accessions representing 12 new sources of EFB resistance were crossed with susceptible selections resulting in 18 seedling populations. Segregation ratios in the seedling populations fit the expected 1:1 ratio for 10 sources, while one source showed an excess of resistant seedlings and another showed an excess of susceptible seedlings. Based on correlation of disease response and scores of SSR markers in the ‘Gasaway’ resistance region in the seedlings, eight resistance sources were assigned to LG6. Linkage maps were constructed for each progeny using SSR markers. The LG6 resistance sources include two selections (#23 and #26) from the Russian Research Institute of Forestry and Mechanization near Moscow, four selections from southern Russia, one selection (OSU 1185.126) from Crimea, one selection (OSU 533.129) from Michigan, Corylus heterophylla ‘Ogyoo’ from the South Korea, and the interspecific hybrid ’Estrella #1’. These new LG6 resistance sources and SSR markers should be useful in breeding new cultivars, including the pyramiding of resistance genes. For the other four resistance sources (Moscow #37, hybrid selection OSU 401.014, C. americana ‘Winkler’ and C. americana OSU 366.060), SSR marker scores on linkage groups 6, 7 and 2 were not correlated with disease response and merit further investigation.

Pyramiding the antimicrobial PR1aCB and AATCB genes in 'Tarocco' blood orange (Citrus sinensis Osbeck) to enhance citrus canker resistance.

Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), is a major bacterial disease responsible for substantial economic losses in citrus-producing areas. To breed transgenic citrus plants with enhanced resistance to citrus canker, two antimicrobial peptide genes, PR1aCB and AATCB, were incorporated into 'Tarocco' blood orange (Citrus sinensis Osbeck) plants via co-transformation and sequential re-transformation. The presence of PR1aCB and AATCB in double transgenic plants was confirmed by PCR. The expression of PR1aCB and AATCB in double transformants was demonstrated by quantitative real-time PCR. An in vivo disease resistance assay involving the injection of Xcc revealed that the double transformants were more resistant to citrus canker than the single gene transformants and wild-type plants. An analysis of the bacterial population indicated that the enhanced citrus canker resistance of the double transformants was due to inhibited Xcc growth. These results proved that the pyramiding of multiple genes is a more effective strategy for increasing the disease resistance of transgenic citrus plants than single gene transformations.

A novel introgression line collection to unravel the genetics of climacteric ripening and fruit quality in melon

Introgression lines are valuable germplasm for scientists and breeders, since they ease genetic studies such as QTL interactions and positional cloning as well as the introduction of favorable alleles into elite varieties. We developed a novel introgression line collection in melon using two commercial European varieties with different ripening behavior, the climacteric cantalupensis ‘Védrantais’ as recurrent parent and the non-climacteric inodorus ‘Piel de Sapo’ as donor parent. The collection contains 34 introgression lines, covering 99% of the donor genome. The mean introgression size is 18.16 Mb and ~ 3 lines were obtained per chromosome, on average. The high segregation of these lines for multiple fruit quality traits allowed us to identify 27 QTLs that modified sugar content, altered fruit morphology or were involved in climacteric ripening. In addition, we confirmed the genomic location of five major genes previously described, which control mainly fruit appearance, such as mottled rind and external color. Most of the QTLs had been reported before in other populations sharing parental lines, while three QTLs (EAROQP11.3, ECDQP11.2 and FIRQP4.1) were newly detected in our work. These introgression lines would be useful to perform additional genetic studies, as fine mapping and gene pyramiding, especially for important complex traits such as fruit weight and climacteric ripening.

Vegetable Breeding Strategies

Variation present in vegetables provides plentiful of opportunity for development of breeding lines and hybrids. Germplasm development depends on incorporation of approaches such as MAGIC population, use of wild relatives, genetic transformation for transferring resistance gene and gene pyramiding. These approaches are most reliable to improve qualitative as well as quantitative characters. Since conventional methods are time consuming, molecular techniques are also utilized at appropriate stages to enhance swiftness of line development. Molecular approaches includes many tools but marker assisted selection is best to use in vegetable breeding programme. It helps to rapid back cross breeding also selection of resistance or qualitative lines from segregating populations. Double haploid is one of finest methods to create diverse pure lines in short interval. There are many ways to incorporate diverse gene in single line through double haploid technology. Hybridization mating such as line x tester or diallel are common methods instead combination breeding method is very result oriented as compared with others. The success rate of specific combination breeding is as much as double as associated with other ones.

Unraveling a genetic roadmap for improved taste in the domesticated apple

Although taste is an important aspect of fruit quality, an understanding of its genetic control remains elusive in apple and other fruit crops. In this study, we conducted genomic sequence analysis of 497 Malus accessions and revealed erosion of genetic diversity caused by apple breeding and possible independent domestication events of dessert and cider apples. Signatures of selection for fruit acidity and size, but not for fruit sugar content, were detected during the processes of both domestication and improvement. Furthermore, we found that single mutations in major genes affecting fruit taste, including Ma1, MdTDT, and MdSOT2, dramatically decrease malate, citrate, and sorbitol accumulation, respectively, and correspond to important domestication events. Interestingly, Ma1 was identified to have pleiotropic effects on both organic acid content and sugar:acid ratio, suggesting that it plays a vital role in determining fruit taste. Fruit taste is unlikely to have been negatively affected by linkage drag associated with selection for larger fruit that resulted from the pyramiding of multiple genes with minor effects on fruit size. Collectively, our study provides new insights into the genetic basis of fruit quality and its evolutionary roadmap during apple domestication, pinpointing several candidate genes for genetic manipulation of fruit taste in apple.

Marker assisted pyramiding of stem rust, leaf rust and powdery mildew resistance genes for durable resistance in wheat (Triticum aestivum L.)

Wheat production is essential for food security. Stem and leaf rust diseases continually pose threat to wheat production. In recent years, climate change and intensive crop cultivation practices are making powdery mildew as a potential threat to wheat production. Deploying resistant cultivars are the most economic, reliable and sustainable way to manage the stem rust, leaf rust and powdery mildew of wheat. Using conventional selection system, it is difficult to select two or more genes in a single genotype. In such a situation, phenotype neutral selection based on marker-trait association along with seedling and adult plant reaction become inevitable. Stem rust, leaf rust and powdery mildew resistance genes, Sr24/Lr24 , Sr26 and Sr36/Pm6 were pyramided in the background of nine Indian wheat cultivars through marker assisted backcross approach. The presence of the rust and powdery mildew resistance genes were confirmed using microsatellite markers such as Sr24#12 ( Sr24/Lr24 ), Sr26#43 ( Sr26 ) and stm773 ( Sr36/Pm6 ) in the pyramided lines. Stable lines were selected at BC 3 F 4 generation. Seedling and adult plant reaction of pyramided lines showed resistance to most of the stem and leaf rust pathotypes prevailing in India. Using gene pyramids ( Sr24/Lr24 , Sr26 and Sr36/Pm6 ) that confer resistance to the predominant pathotypes of stem rust, leaf rust and powdery mildew could impart durability to the cultivars than single gene deployment.

Virulence structure of Blumeria graminis f. sp. tritici, the causal agent of wheat powdery mildew, in Ontario, Canada, in 2018 and 2019

Powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt) is a major disease of wheat (Triticum aestivum) in Ontario, which can cause 20% yield loss. The development of resistant commercial wheat cultivars is the most economical means of controlling this disease, but only if the resistance genes used are incompatible with the virulence phenotypes present in the pathogen population. The virulence structure of Bgt in Ontario was examined in 2018 and 2019. Of the 42 single colony isolates collected in Ontario greenhouses and commercial fields, 40 virulence phenotypes, assigned VP1 to VP40, were identified on a set of 24 single-gene differential genotypes. Of the 24 resistance genes possessed by the differential genotypes, eight genes including Pm1a, Pm1b, Pm1c, Pm12, Pm16, Pm21, Pm37, and MlAG12 were effective against all of the Bgt isolates. Four genes including Pm3d, Pm29, Pm34, and NCAG13 were mostly effective with resistance reactions to more than 80% of the isolates. There were no significant differences in numbers of virulence genes per isolate between the two years, or between the greenhouse and field origins. The virulence frequencies of Bgt isolates for these effective and mostly effective genes were also not significantly affected by the year of collection and their origins, suggesting that the Bgt population is relatively stable. The effective genes identified in this study may be deployed singly or used for gene pyramiding in wheat breeding programs for developing powdery mildew-resistant cultivars in Ontario.

Molecular and phenotypic evaluation for heading date and panicle length on F2 selected aromatic rice progenies derived from gene pyramiding effort

Gene pyramiding, an attempt to combine many favourable genes from many parents into one single genotype, has been applied in recent years. Through hybridization of many rice genotypes including cv. Pandanwangi (fragrance rice), PTB33 (brown planthopper resistant), Ciapus (high yield and moderate amylose content) and KA (early maturity), some promising genotypes have been obtained. Thus, in order to select the best genotypes among the F2 progenies derived from gene pyramiding effort, both phenotypic and molecular evaluations have to be done. These pyramided genotypes were evaluated with IFAP and ESP (fgr gene), RM19414 (Hd3), RM7601 (Hd2), and RM3600 (LP1) for molecular assessment, meanwhile morpho-agronomic traits were done to assess their performance. Aromatic trait was detected by 1.7% KOH. Meanwhile heading date and panicle length were measured phenotypically. Data obtained from molecular markers were scanned by visualizing DNA bands and calculating PIC (Polymorphism Information Content). Data from phenotypic markers were categorized based on IBPGR (International Board for Plant Genetic Resources) standards. Genotype #131 was supposed to have high yield potential, early maturity, aromatic, and intermediate amylose content. The selected will be recommended as promising genotype and continued as elite breeding materials and will be evaluated for their benefit.