Tissue culture and Agrobacterium-mediated genetic transformation of the oil crop sunflower.

Sunflower is one of the four major oil crops in the world. 'Zaoaidatou' (ZADT), the main variety of oil sunflower in the northwest of China, has a short growth cycle, high yield, and high resistance to abiotic stress. However, the ability to tolerate adervesity is limited. Therefore, in this study, we used the retention line of backbone parent ZADT as material to establish its tissue culture and genetic transformation system for new variety cultivating to enhance resistance and yields by molecular breeding. The combination of 0.05 mg/L IAA and 2 mg/L KT in MS was more suitable for direct induction of adventitious buds with cotyledon nodes and the addition of 0.9 mg/L IBA to MS was for adventitious rooting. On this basis, an efficient Agrobacterium tumefaciens-mediated genetic transformation system for ZADT was developed by the screening of kanamycin and optimization of transformation conditions. The rate of positive seedlings reached 8.0%, as determined by polymerase chain reaction (PCR), under the condition of 45 mg/L kanamycin, bacterial density of OD600 0.8, infection time of 30 min, and co-cultivation of three days. These efficient regeneration and genetic transformation platforms are very useful for accelerating the molecular breeding process on sunflower.

Identification of Induced Copy Number Variation from Low Coverage Sequence Data.

Induced mutations have been an important tool for plant breeding and functional genomics for more than 80 years. Novel mutations can be induced by treating seed or other plant cells with chemical mutagens or ionizing radiation. The majority of released mutant crop varieties were developed using ionizing radiation. This has been shown to create a variety of different DNA lesions including large (e.g., >=10,000 bps) copy number variations (CNV). Detection of induced DNA lesions from whole genome sequence data is useful for choosing a mutagen dosage prior to committing resources to develop a large mutant population for forward or reverse-genetic screening. Here I provide a method for detecting large induced CNV from mutant plants that utilizes a new tool to streamline the process of obtaining read coverage directly from BAM files, comparing non-mutagenized controls and mutagenized samples, and plotting the results for visual evaluation. Example data is provided from low coverage sequence data from gamma-irradiated vegetatively propagated triploid banana.

Breeding of ornamental orchids with focus on Phalaenopsis: current approaches, tools, and challenges for this century.

Ornamental orchid breeding programs have been conducted to develop commercially valuable cultivars with improved characteristics of commercial interest, such as size, flower color, pattern, shape, and resistance to pathogens. Conventional breeding, including sexual hybridization followed by selection of desirable characteristics in plants, has so far been the main method for ornamental breeding, but other techniques, including mutation induction by polyploidization and gamma irradiation, and biotechnological techniques, such as genetic transformation, have also been studied and used in ornamental breeding programs. Orchids are one of the most commercially important families in floriculture industry, having very particular reproductive biology characteristics and being a well-studied group of ornamentals in terms of genetic improvement. The present review focuses on the conventional and biotechnological techniques and approaches specially employed in breeding Phalaenopsis orchids, the genus with highest worldwide importance as an ornamental orchid, highlighting the main limitations and strengths of the approaches. Furthermore, new opportunities and future prospects for ornamental breeding in the CRISPR/Cas9 genome editing era are also discussed. We conclude that conventional hybridization remains the most used method to obtain new cultivars in orchids. However, the emergence of the first biotechnology-derived cultivars, as well as the new biotechnological tools available, such as CRISPR-Cas9, rekindled the full potential of biotechnology approaches and their importance for improve ornamental orchid breeding programs.

Genotyping by sequencing for the construction of oil palm (Elaeis guineensis Jacq.) genetic linkage map and mapping of yield related quantitative trait loci.

Background: Oil palm (Elaeis guineensis Jacq.) is one of the major oil-producing crops. Improving the quality and increasing the production yield of oil palm have been the primary focuses of both conventional and modern breeding approaches. However, the conventional breeding approach for oil palm is very challenging due to its longevity, which results in a long breeding cycle. Thus, the establishment of marker assisted selection (MAS) for oil palm breeding programs would speed up the breeding pipeline by generating new oil palm varieties that possess high commercial traits. With the decreasing cost of sequencing, Genotyping-by-sequencing (GBS) is currently feasible to many researchers and it provides a platform to accelerate the discovery of single nucleotide polymorphism (SNP) as well as insertion and deletion (InDel) markers for the construction of a genetic linkage map. A genetic linkage map facilitates the identification of significant DNA regions associated with the trait of interest via quantitative trait loci (QTL) analysis. Methods: A mapping population of 112 F1 individuals from a cross of Deli dura and Serdang pisifera was used in this study. GBS libraries were constructed using the double digestion method with HindIII and TaqI enzymes. Reduced representation libraries (RRL) of 112 F1 progeny and their parents were sequenced and the reads were mapped against the E. guineensis reference genome. To construct the oil palm genetic linkage map, informative SNP and InDel markers were used to discover significant DNA regions associated with the traits of interest. The nine traits of interest in this study were fresh fruit bunch (FFB) yield, oil yield (OY), oil to bunch ratio (O/B), oil to dry mesocarp ratio (O/DM) ratio, oil to wet mesocarp ratio (O/WM), mesocarp to fruit ratio (M/F), kernel to fruit ratio (K/F), shell to fruit ratio (S/F), and fruit to bunch ratio (F/B). Results: A total of 2.5 million SNP and 153,547 InDel markers were identified. However, only a subset of 5,278 markers comprising of 4,838 SNPs and 440 InDels were informative for the construction of a genetic linkage map. Sixteen linkage groups were produced, spanning 2,737.6 cM for the maternal map and 4,571.6 cM for the paternal map, with average marker densities of one marker per 2.9 cM and one per 2.0 cM respectively, were produced. A QTL analysis was performed on nine traits; however, only QTL regions linked to M/F, K/F and S/F were declared to be significant. Of those QTLs were detected: two for M/F, four for K/F and one for S/F. These QTLs explained 18.1-25.6% of the phenotypic variance and were located near putative genes, such as casein kinase II and the zinc finger CCCH domain, which are involved in seed germination and growth. The identified QTL regions for M/F, K/F and S/F from this study could be applied in an oil palm breeding program and used to screen palms with desired traits via marker assisted selection (MAS).

Metabolic profiles of peanut (Arachis hypogaea L.) in response to Puccinia arachidis fungal infection.

Background Puccinia arachidis fungus causes rust disease in the peanut plants (Arachis hypogaea L.), which leads to high yield loss. Metabolomic profiling of Arachis hypogaea was performed to identify the pathogen-induced production of metabolites involved in the defense mechanism of peanut plants. In this study, two peanut genotypes, one susceptible (JL-24) and one resistant (GPBD-4) were inoculated with Puccinia arachidis fungal pathogen. The metabolic response was assessed at the control stage (0 day without inoculation), 2 DAI (Day after inoculation), 4 DAI and 6 DAI by Gas Chromatography-Mass Spectrometry (GC-MS). Results About 61 metabolites were identified by NIST library, comprising sugars, phenols, fatty acids, carboxylic acids and sugar alcohols. Sugars and fatty acids were predominant in leaf extracts compared to other metabolites. Concentration of different metabolites such as salicylic acid, mannitol, flavonoid, 9,12-octadecadienoic acid, linolenic acid and glucopyranoside were higher in resistant genotype than in susceptible genotype during infection. Systemic acquired resistance (SAR) and hypersensitive reaction (HR) components such as oxalic acid was elevated in resistant genotype during pathogen infection. Partial least square-discriminant analysis (PLS-DA) was applied to GC-MS data for revealing metabolites profile between resistant and susceptible genotype during infection. Conclusion The phenol content and oxidative enzyme activity i.e. catalase, peroxidase and polyphenol oxidase were found to be very high at 4 DAI in resistant genotype (p-value < 0.01). This metabolic approach provides information about bioactive plant metabolites and their application in crop protection and marker-assisted plant breeding.

The RUBY reporter enables efficient haploid identification in maize and tomato.

In vivo haploid induction has been extended from maize to monocotyledonous plants like rice, wheat, millet and dicotyledonous plants such as tomato, rapeseed, tobacco and cabbage. Accurate identification of haploids is a crucial step of doubled haploid technology, where a useful identification marker is very pivotal. R1-nj is an extensively used visual marker for haploid identification in maize. RFP and eGFP have been shown to be feasible in identifying haploid. However, these methods are either limited to specific species, or require specific equipment. It still lacks an efficient visual marker that is practical across different crop species. In this study, we introduced the RUBY reporter, a betalain biosynthesis system, into maize and tomato haploid inducers as a new marker for haploid identification. Results showed that expression of RUBY could result in deep betalain pigmentation in maize embryos as early as 10 days after pollination, and enabled 100% accuracy of immature haploid embryo identification. Further investigation in tomato revealed that the new marker led to deep red pigmentation in radicles and haploids can be identified easily and accurately. The results demonstrated that the RUBY reporter is a background-independent and efficient marker for haploid identification and would be promising in doubled haploid breeding across different crop species.

Inheritance of seedlessness and the molecular characterization of the INO gene in Annonaceae / Herança da ausência de sementes e a caracterização molecular do gene INO em Annonaceae

The inheritance of the seedless fruit characteristic of Annona squamosa has not yet been explained. Molecular techniques may aid breeding programs, mainly in the assisted selection of the target gene. The INO gene may be related to seed development in these fruits. The objective of the present paper was to investigate the inheritance of seedlessness in the 'Brazilian seedless' sugar apple and INO gene conservation in Annona squamosa and Annona cherimola x Annona squamosa genotypes by assessing their homology with the INO database genes. The F1 generation was obtained by crossing the mutant 'Brazilian seedless' (male genitor) (P1) with the wild-type A. squamosa with seeds (M1 and M2, female genitors). The INO gene was studied in mutant and wild-type A. squamosa (P1, M1, M2 and M3) and in the Gefner atemoya (A. cherimola x A. squamosa) (M4) cultivar. The DNA was extracted from young leaves, and four sets of specific primers flanking the INO gene were amplified. The seedless characteristic was identified as stenospermatic in the fruits of parental P1, suggesting monogenic inheritance with complete dominance. High sequence similarity of the INO gene amplifications in the sugar apple accessions (M1, M2, M3) and the atemoya cultivar Gefner (M4) reinforces the hypothesis of their conservation.

A herança da característica de fruto sem sementes de Annona squamosa ainda não foi esclarecida. Técnicas moleculares podem auxiliar em programas de melhoramento, principalmente na seleção assistida do gene de interesse. O gene INO pode estar relacionado ao desenvolvimento da semente dessas frutas. O objetivo foi investigar a herança da ausência de sementes em Annona squamosa e a conservação do gene INO nos genótipos Annona squamosa e Annona cherimola x Annona squamosa avaliando sua homologia com banco de dados de genes INO. A geração F1 foi obtida pelo cruzamento do mutante 'Brazilian seedless' (genitor masculino) (P1) com o tipo selvagem com sementes (A. squamosa) (M1 e M2, genitores femininos). O gene INO foi estudado em A. squamosa, mutante e selvagem (P1, M1, M2 e M3) e na cultivar Gefner atemoya (A. cherimola x A. squamosa) (M4). O DNA foi extraído de folhas jovens, e quatro conjuntos de primers específicos flanqueando o gene INO foram amplificados. A característica sem sementes foi identificada como estenospermática nos frutos do parental P1, sugerindo herança monogênica com dominância completa. A alta similaridade de sequência das amplificações do gene INO nos acessos de pinha (M1, M2, M3) e na cultivar de atemóia Gefner (M4) reforça a hipótese de sua conservação.

Characterization of wheat-Thinopyrum bessarabicum genetic stock for stripe rust and Karnal bunt resistance / Caracterização do estoque genético de trigo-Thinopyrum bessarabicum para resistência de ferrugem e Karnal bunt

Utilization of modern breeding techniques for developing high yielding and uniform plant types ultimately narrowing the genetic makeup of most crops. Narrowed genetic makeup of these crops has made them vulnerable towards disease and insect epidemics. For sustainable crop production, genetic variability of these crops must be broadened against various biotic and abiotic stresses. One of the ways to widen genetic configuration of these crops is to identify novel additional sources of durable resistance. In this regard crops wild relatives are providing valuable sources of allelic diversity towards various biotic, abiotic stress tolerance and quality components. For incorporating novel variability from wild relative’s wide hybridization technique has become a promising breeding method. For this purpose, wheat-Th. bessarabicum amphiploid, addition and translocation lines have been screened in field and screen house conditions to get novel sources of yellow rust and Karnal bunt resistant. Stripe rust screening under field conditions has revealed addition lines 4JJ and 6JJ as resistant to moderately resistant while addition lines 3JJ, 5JJ, 7JJ and translocation lines Tr-3, Tr-6 as moderately resistant wheat-Thinopyrum-bessarabicum genetic stock. Karnal bunt screening depicted addition lines 5JJ and 4JJ as highly resistant genetic stock. These genetic stocks may be used to introgression novel stripe rust and Karnal bunt resistance from the tertiary gene pool into susceptible wheat backgrounds.

A utilização de técnicas modernas de melhoramento para o desenvolvimento de tipos de plantas uniformes e de alto rendimento, em última análise, estreitando a composição genética da maioria das culturas. A composição genética restrita dessas plantações tornou-as vulneráveis a doenças e epidemias de insetos. Para uma produção agrícola sustentável, a variabilidade genética dessas culturas deve ser ampliada contra vários estresses bióticos e abióticos. Uma das maneiras de ampliar a configuração genética dessas culturas é identificar novas fontes adicionais de resistência durável. A esse respeito, os parentes selvagens das culturas estão fornecendo fontes valiosas de diversidade alélica para vários componentes de qualidade e tolerância ao estresse abiótico e biótico. Para incorporar a nova variabilidade da ampla técnica de hibridização de parente selvagem tornou-se um método de reprodução promissor. Para esse efeito, trigo-Th. As linhas anfiploides, de adição e translocação de bessarabicum foram selecionadas em condições de campo e de casa de tela para obter novas fontes de ferrugem amarela e resistência ao bunt de Karnal. A triagem de ferrugem em faixas em condições de campo revelou as linhas de adição 4JJ e 6JJ como resistentes a moderadamente resistentes, enquanto as linhas de adição 3JJ, 5JJ, 7JJ e as linhas de translocação Tr-3, Tr-6 como estoque genético de trigo-Thinopyrum bessarabicum moderadamente resistente. A triagem Karnal bunt descreveu as linhas de adição 5JJ e 4JJ como estoque genético altamente resistente. Esses estoques genéticos podem ser usados para introgressão da nova ferrugem e resistência ao bunt de Karnal do pool genético terciário em origens de trigo suscetíveis.

Advances in protoplast transfection promote efficient CRISPR/Cas9-mediated genome editing in tetraploid potato.

MAIN CONCLUSION: An efficient method of DNA-free gene-editing in potato protoplasts was developed using linearized DNA fragments, UBIQUITIN10 promoters of several plant species, kanamycin selection, and transient overexpression of the BABYBOOM transcription factor. Plant protoplasts represent a reliable experimental system for the genetic manipulation of desired traits using gene editing. Nevertheless, the selection and regeneration of mutated protoplasts are challenging and subsequent recovery of successfully edited plants is a significant bottleneck in advanced plant breeding technologies. In an effort to alleviate the obstacles related to protoplasts' transgene expression and protoplasts' regeneration, a new method was developed. In so doing, it was shown that linearized DNA could efficiently transfect potato protoplasts and that UBIQUITIN10 promoters from various plants could direct transgene expression in an effective manner. Also, the inhibitory concentration of kanamycin was standardized for transfected protoplasts, and the NEOMYCIN PHOSPHOTRANSFERASE2 (NPT2) gene could be used as a potent selection marker for the enrichment of transfected protoplasts. Furthermore, transient expression of the BABYBOOM (BBM) transcription factor promoted the regeneration of protoplast-derived calli. Together, these methods significantly increased the selection for protoplasts that displayed high transgene expression, and thereby significantly increased the rate of gene editing events in protoplast-derived calli to 95%. The method developed in this study facilitated gene-editing in tetraploid potato plants and opened the way to sophisticated genetic manipulation in polyploid organisms.

Silicon-enhanced tolerance to cadmium toxicity in soybean by enhancing antioxidant defense capacity and changing cadmium distribution and transport.

Cadmium (Cd) is a widely distributed heavy metal that is toxic to plants and humans. Although silicon (Si) has been reported to reduce Cd accumulation and toxicity in plants, evidence on the functions of Si and its mechanisms in the possible alleviation of soybean are limited. Therefore, a controlled experiment was conducted to investigate the impacts and mechanisms of Si on Cd retention in soybean. Here, we determined the growth index, Cd distribution, and antioxidant activity systems of Si, as well as expression levels of differentially expressed genes (DEGs) in Si under Cd stress, and conducted RNA-seq analysis. We not only found that Si can significantly promote soybean plant growth, increase plant antioxidant activities, and reduce the Cd translocation factor, but also revealed that a total of 636 DEGs were shared between CK and Cd, CK and Cd + Si, and Cd and Cd + Si. Moreover, several genes were significantly enriched in antioxidant systems and Cd distribution and transport systems. Therefore, the expression status of Si-mediated Cd stress response genes is likely involved in improving oxidative stress and changing Cd uptake and transport, as well as improving plant growth that contributes to Si alleviating Cd toxicity in plants. Moreover, numerous potential target genes were identified for the engineering of Cd-tolerant cultivars in soybean breeding programs.

Comparative Cytological and Transcriptome Analyses of Anther Development in Nsa Cytoplasmic Male Sterile (1258A) and Maintainer Lines in Brassica napus Produced by Distant Hybridization.

1258A is a new line of B.napus with Nsa cytoplasmic male sterility (CMS) with potential applications in hybrid rapeseed breeding. Sterile cytoplasm was obtained from XinJiang Sinapis arvensis through distant hybridization and then backcrossed with 1258B for many generations. However, the characteristics and molecular mechanisms underlying pollen abortion in this sterile line are poorly understood. In this study, a cytological analysis revealed normal microsporogenesis and uninucleate pollen grain formation. Pollen abortion was due to non-programmed cell death in the tapetum and the inability of microspores to develop into mature pollen grains. Sucrose, soluble sugar, and adenosine triphosphate (ATP) contents during microspore development were lower than those of the maintainer line, along with an insufficient energy supply, reduced antioxidant enzyme activity, and substantial malondialdehyde (MDA) accumulation in the anthers. Transcriptome analysis revealed that genes involved in secondary metabolite biosynthesis, glutathione metabolism, phenylpropane biosynthesis, cyanoamino acid metabolism, starch and sucrose metabolism, and glycerolipid metabolism may contribute to pollen abortion. The down regulation of nine cytochrome P450 monooxygenases genes were closely associated with pollen abortion. These results suggest that pollen abortion in 1258A CMS stems from abnormalities in the chorioallantoic membranes, energy deficiencies, and dysfunctional antioxidant systems in the anthers. Our results provide insight into the molecular mechanism underlying pollen abortion in Nsa CMS and provide a theoretical basis for better heterosis utilization in B.napus.

A transcriptome-based association study of growth, wood quality, and oleoresin traits in a slash pine breeding population.

Slash pine (Pinus elliottii Engelm.) is an important timber and resin species in the United States, China, Brazil and other countries. Understanding the genetic basis of these traits will accelerate its breeding progress. We carried out a genome-wide association study (GWAS), transcriptome-wide association study (TWAS) and weighted gene co-expression network analysis (WGCNA) for growth, wood quality, and oleoresin traits using 240 unrelated individuals from a Chinese slash pine breeding population. We developed high quality 53,229 single nucleotide polymorphisms (SNPs). Our analysis reveals three main results: (1) the Chinese breeding population can be divided into three genetic groups with a mean inbreeding coefficient of 0.137; (2) 32 SNPs significantly were associated with growth and oleoresin traits, accounting for the phenotypic variance ranging from 12.3% to 21.8% and from 10.6% to 16.7%, respectively; and (3) six genes encoding PeTLP, PeAP2/ERF, PePUP9, PeSLP, PeHSP, and PeOCT1 proteins were identified and validated by quantitative real time polymerase chain reaction for their association with growth and oleoresin traits. These results could be useful for tree breeding and functional studies in advanced slash pine breeding program.

BnERF114.A1, a Rapeseed Gene Encoding APETALA2/ETHYLENE RESPONSE FACTOR, Regulates Plant Architecture through Auxin Accumulation in the Apex in Arabidopsis.

Plant architecture is crucial for rapeseed breeding. Here, we demonstrate the involvement of BnERF114.A1, a transcription factor for ETHYLENE RESPONSE FACTOR (ERF), in the regulation of plant architecture in Brassica napus. BnERF114.A1 is a member of the ERF family group X-a, encoding a putative 252-amino acid (aa) protein, which harbours the AP2/ERF domain and the conserved CMX-1 motif. BnERF114.A1 is localised to the nucleus and presents transcriptional activity, with the functional region located at 142-252 aa of the C-terminus. GUS staining revealed high BnERF114.A1 expression in leaf primordia, shoot apical meristem, leaf marginal meristem, and reproductive organs. Ectopic BnERF114.A1 expression in Arabidopsis reduced plant height, increased branch and silique number per plant, and improved seed yield per plant. Furthermore, in Arabidopsis, BnERF114.A1 overexpression inhibited indole-3-acetic acid (IAA) efflux, thus promoting auxin accumulation in the apex and arresting apical dominance. Therefore, BnERF114.A1 probably plays an important role in auxin-dependent plant architecture regulation.

Random mutagenesis in vegetatively propagated crops: opportunities, challenges and genome editing prospects.

In order to meet the growing human food and nutrition demand a perpetual process of crop improvement is idealized. It has seen changing trends and varying concepts throughout human history; from simple selection to complex gene-editing. Among these techniques, random mutagenesis has been shown to be a promising technology to achieve desirable genetic gain with less time and minimal efforts. Over the decade, several hundred varieties have been released through random mutagenesis, but the production is falling behind the demand. Several food crops like banana, potato, cassava, sweet potato, apple, citrus, and others are vegetatively propagated. Since such crops are not propagated through seed, genetic improvement through classical breeding is impractical for them. Besides, in the case of polyploids, accomplishment of allelic homozygosity requires a considerable land area, extensive fieldwork with huge manpower, and hefty funding for an extended period of time. Apart from induction, mapping of induced genes to facilitate the knowledge of biological processes has been performed only in a few selected facultative vegetative crops like banana and cassava which can form a segregating population. During the last few decades, there has been a shift in the techniques used for crop improvement. With the introduction of the robust technologies like meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats (CRISPR) more and more crops are being subjected to gene editing. However, more work needs to be done in case of vegetatively propagated crops.

Integrating omics approaches to discover and prioritize candidate genes involved in oil biosynthesis in soybean.

Soybean is a major source of edible protein and oil. Oil content is a quantitative trait that is significantly determined by genetic and environmental factors. Over the past 30 years, a large volume of soybean genetic, genomic, and transcriptomic data have been accumulated. Nevertheless, integrative analyses of such data remain scarce, in spite of their importance for crop improvement. We hypothesized that the co-occurrence of genomic regions for oil-related traits in different studies may reveal more stable regions encompassing important genetic determinants of oil content and quality in soybean. We integrated publicly available data, obtained with distinct techniques, to discover and prioritize candidate genes involved in oil biosynthesis and regulation in soybean. We detected key fatty acid biosynthesis genes (e.g., BCCP2 and ACCase, FADs, KAS family proteins) and several transcription factors, which are likely regulators of oil biosynthesis. In addition, we identified new candidates for seed oil accumulation and quality, such as Glyma.03G213300 and Glyma.19G160700, which encode a translocator protein homolog and a histone acetyltransferase, respectively. Further, oil and protein genomic hotspots are strongly associated with breeding and not with domestication, suggesting that soybean domestication prioritized other traits. The genes identified here are promising targets for breeding programs and for the development of soybean lines with increased oil content and quality.

Genetic parameters controlling the inheritance of glaucousness and yield traits in bread wheat / Parâmetros genéticos que controlam a herança de glaucosidade e características de produção em trigo mole

Wheat breeders frequently use generation mean analysis to obtain information on the type of gene action involved in inheriting a trait to choose the helpful breeding procedure for trait improvement. The present study was carried out to study the inter-allelic and intra-allelic gene action and inheritance of glaucousness, earliness and yield traits in a bread wheat cross between divergent parents in glaucousness and yield traits; namely Mut-2 (P1) and Sakha 93 (P2). The experimental material included six populations, i.e. P1, P2, F1, F2, BC1, and BC2 for this wheat cross. A randomized complete block design with three replications was used, and a six parameters model was applied. Additive effects were generally more critical than dominance for all studied traits, except for plant height (PH) and grain yield/plant (GYPP). The duplicate epistasis was observed in spike length; SL, spikes/plant; SPP and days to heading; DTH. All six types of allelic and non-allelic interaction effects controlled SL, GYPP, DTH and glaucousness. All three types of epistasis, i.e. additive x additive, additive x dominance, and dominance x dominance, are essential in determining the inheritance of four traits (SL, GYPP, DTH and glaucousness). Dominance × dominance effects were higher in magnitude than additive × dominance and additive × additive in most traits. The average degree of dominance was minor than unity in six traits (glaucousness, grains/spike, spike weight, days to maturity, 100-grain weight and SL), indicating partial dominance and selection for these traits might be more effective in early generations. Meanwhile, the remaining traits (PH, SPP, GYPP and DTH) had a degree of dominance more than unity, indicating that overdominance gene effects control such traits and it is preferable to postpone selection to later generations. The highest values of [...].

Os criadores de trigo frequentemente usam a análise da média de geração para obter informações sobre o tipo de ação do gene envolvida na herança de uma característica para escolher o procedimento de melhoramento útil para o aprimoramento da característica. O presente estudo foi conduzido para estudar a ação do gene interalélico e intraalélico e a herança de características de glaucosidade, precocidade e produção em um cruzamento de trigo mole entre pais divergentes em glaucosidade e características de produção; nomeadamente Mut-2 (P1) e Sakha 93 (P2). O material experimental incluiu seis populações, ou seja, P1, P2, F1, F2, BC1 e BC2 para este cruzamento de trigo. O delineamento experimental foi em blocos ao acaso com três repetições e aplicado um modelo de seis parâmetros. Os efeitos aditivos foram geralmente mais críticos do que a dominância para todas as características estudadas, exceto para altura da planta (AP) e rendimento de grãos / planta (GYPP). A epistasia duplicada foi observada no comprimento da ponta; SL, espigas / planta; SPP e dias para o cabeçalho; DTH. Todos os seis tipos de efeitos de interação alélica e não alélica controlaram SL, GYPP, DTH e glaucosidade. Todos os três tipos de epistasia, ou seja, aditivo x aditivo, aditivo x dominância e dominância x dominância, são essenciais na determinação da herança de quatro características (SL, GYPP, DTH e glaucosidade). Os efeitos de dominância × dominância foram maiores em magnitude do que aditivo × dominância e aditivo × aditivo na maioria das características. O grau médio de dominância foi menor do que a unidade em seis características (glaucosidade, grãos / espiga, peso da espiga, dias até a maturidade, peso de 100 grãos e SL), indicando dominância parcial, e a seleção para essas características pode ser mais eficaz nas gerações iniciais. Enquanto isso, os traços restantes (PH, SPP, GYPP e [...].

Genotype plus genotype by-environment interaction biplot and genetic diversity analyses on multi-environment trials data of yield and technological traits of cotton cultivars / Biplot de genótipo mais interação genótipo por ambiente e diversidade genética em dados de produtividade e qualidade de fibras em cultivares de algodão provenientes de ensaios multi-ambientes

Understanding the genetic diversity and overcoming genotype-by-environment interaction issues is an essential step in breeding programs that aims to improve the performance of desirable traits. This study estimated genetic diversity and applied genotype + genotype-by-environment (GGE) biplot analyses in cotton genotypes. Twelve genotypes were evaluated for fiber yield, fiber length, fiber strength, and micronaire. Estimation of variance components and genetic parameters was made through restricted maximum likelihood and the prediction of genotypic values was made through best linear unbiased prediction. The modified Tocher and principal component analysis (PCA) methods, were used to quantify genetic diversity among genotypes. GGE biplot was performed to find the best genotypes regarding adaptability and stability. The Tocher technique and PCA allowed for the formation of clusters of similar genotypes based on a multivariate framework. The GGE biplot indicated that the genotypes IMACV 690 and IMA08 WS were highly adaptable and stable for the main traits in cotton. The cross between the genotype IMACV 690 and IMA08 WS is the most recommended to increase the performance of the main traits in cotton crops.

Compreender a diversidade genética e contornar os problemas causados pela interação genótipos por ambientes é uma etapa importante em programas de melhoramento. Este estudo teve como objetivo estimar a diversidade genética e aplicar a metodologia de biplot genótipo + genótipo por ambiente (GGE biplot) em doze genótipos de algodão avaliados quanto ao rendimento da fibra, comprimento da fibra, resistência da fibra e micronaire. A estimativa dos componentes de variância e dos parâmetros genéticos foi feita através do método da máxima verossimilhança restrita e a predição dos valores genotípicos por meio da melhor predição linear não enviesada. Os métodos de Tocher modificado e análise de componentes principais (PCA) foram utilizados para quantificar a diversidade genética entre os genótipos. O método GGE biplot foi conduzido para encontrar os melhores genótipos em relação à adaptabilidade e estabilidade. As técnicas de Tocher e PCA permitiram a formação de clusters de genótipos semelhantes com base em uma estrutura multivariada. O GGE biplot indicou que os genótipos IMACV 690 e IMA08 WS foram altamente adaptáveis e estáveis para as principais características do algodão. O cruzamento dentre os genótipos IMACV 690 e IMA08 WS é o mais recomendado para aumentar o desempenho das principais características na cultura do algodão.

QTL mapping in outbred tetraploid (and diploid) diallel populations.

Over the last decade, multiparental populations have become a mainstay of genetics research in diploid species. Our goal was to extend this paradigm to autotetraploids by developing software for quantitative trait locus (QTL) mapping in connected F1 populations derived from a set of shared parents. For QTL discovery, phenotypes are regressed on the dosage of parental haplotypes to estimate additive effects. Statistical properties of the model were explored by simulating half-diallel diploid and tetraploid populations with different population sizes and numbers of parents. Across scenarios, the number of progeny per parental haplotype (pph) largely determined the statistical power for QTL detection and accuracy of the estimated haplotype effects. Multiallelic QTL with heritability 0.2 were detected with 90% probability at 25 pph and genome-wide significance level 0.05, and the additive haplotype effects were estimated with over 90% accuracy. Following QTL discovery, the software enables a comparison of models with multiple QTL and nonadditive effects. To illustrate, we analyzed potato tuber shape in a half-diallel population with three tetraploid parents. A well-known QTL on chromosome 10 was detected, for which the inclusion of digenic dominance lowered the Deviance Information Criterion (DIC) by 17 points compared to the additive model. The final model also contained a minor QTL on chromosome 1, but higher-order dominance and epistatic effects were excluded based on the DIC. In terms of practical impacts, the software is already being used to select offspring based on the effect and dosage of particular haplotypes in breeding programs.

Repairing TALEN-mediated double-strand break by microhomology-mediated recombination in tobacco plastids generates abundant subgenomic DNA.

Transcription activator-like effector nuclease (TALEN) technology has been widely used to edit nuclear genomes in plants but rarely for editing organellar genomes. In addition, ciprofloxacin, commonly used to cause the double-strand break of organellar DNA for studying the repair mechanism in plants, confers no organellar selectivity and site-specificity. To demonstrate the feasibility of TALEN-mediated chloroplast DNA editing and to use it for studying the repair mechanism in plastids, we developed a TALEN-mediated editing technology fused with chloroplast transit peptide (cpTALEN) to site-specifically edit the rpoB gene via Agrobacteria-mediated transformation of tobacco leaf. Transgenic plants showed various degrees of chlorotic phenotype. Repairing damaged plastid DNA resulted in point mutation, large deletion and small inversion surrounding the rpoB gene by homologous recombination and/or microhomology-mediated recombination. In an albino line, microhomology-mediated recombination via a pair of 12-bp direct repeats between rpoC2 and ycf2 genes generated the chimeric ycf2-rpoC2 subgenome, with the level about 3- to 5-fold higher for subgenomic DNA than ycf2. Additionally, the expression of chimeric ycf2-rpoC2 transcripts versus ycf2 mRNA agreed well with the level of corresponding DNA. The ycf2-rpoC2 subgenomic DNA might independently and preferentially replicate in plastids.