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Diallel analysis of production traits among domestic, exotic and mutant germplasms of Lycopersicon.

The effects of wild germplasm on tomato fruit shelf life have not yet been completely evaluated. Three different genotypes of Lycopersicon esculentum (a cultivated variety, a homozygote for nor and a homozygote for rin), LA1385 of L. esculentum var. cerasiforme, LA722 of L. pimpinellifolium, and 10 diallel hybrids were assayed. Mean values of fruit shelf life, weight, shape, and mean number of flowers per cluster were analyzed after Griffing (1956, Aust. J. Biology 9: 463-493), method 2, model 1. Both general and specific combining abilities (GCA and SCA) were significant for the four traits. Negative unidirectional dominance was detected for fruit weight and shelf life, while bidirectional dominance was detected for fruit shape and mean number of flowers per cluster. SCA was greater than GCA for shelf life, so nonadditive effects predominantly accounted for this trait. In the heterozygous state, rin had smaller mean effects than nor. Wild accessions were able to prolong shelf life per se, and in crosses to the cultivated variety. The cross between the homozygote for nor and LA722 yielded the longest shelf life among hybrids.

Crosses, Genetic↗

[Preservation of germplasm of Rehmannia glutinosa in vitro].

OBJECTIVE: To establish a method for the germplasm preservation of R. glutinosa. METHOD: Plantlets of different cultivars obtained by tip culture were inoculated into test tubes with MS medium supplemented with 0.2 mg.L-1 BA and 0.02 mg.L-1 NAA, and preserved at 4-6 degrees C in dark. At the same time, different media (A. distilled water + 10 g.L-1 agar; B. 1/2 MS + 5 g.L-1 agar; C. MS + 10 g.L-1 agar; D. 1/2 MS + 0.5 BA + 0.02 NAA + 10 g.L-1 agar; E. MS + 0.5 BA + 0.02 NAA + 10 g.L-1 agar) were set to conserve plantlets of "85-5" on the same condition. RESULT: 4 months later, the death rate of "Xinggeda" was 73%, "Tucheng" 60%, "85-5" 33%, and "Beijing 1" 9%. All of the plantlets of "85-5" in different media kept alive. 10 months later, most of the preserved plantlets browned and wilted except those on medium A. CONCLUSION: The low temperature endurance of R. glutinosa is cultivar-dependent. Medium A can preserve "85-5" for more than 10 months in vitro.

Agar↗

[Analysis of yellow peach germplasm revealed by RAPD markers].

Genomic DNAs of 37 cultivars of yellow peach (Amgdalus persica) were amplified by RAPD (Randomly Amplified Polymorphic DNA) with 22 primers selected from 200 arbitrary 10 decamer primers. Phylogenetic relationship was analyzed through cluster of bands in 180 amplified sites. Molecular checking index and conservation of key germplasm were put forward according to special bands.

English Abstract↗

[Identification and molecular cytology analysis of novel wheat germplasm expressing seven high molecular weight glutenin subunits].

This paper describes the characterization of novel wheat lines NR98116-9-2 and NR98116-9-3 derived from crosses of wheat with hexaploid triticale and expressing seven different high molecular weight glutenin subunits by SDS-PAGE analysis. The results suggested that the two lines were similar to wheat in morphological characters and were stable genetically. The chromosome number of root tip cells of the two lines were 2n=42 and 2n=44 and the configuration of the pollen mother cells at metaphase I were 21 // and 22 //, respectively. The analysis of chromosome constitution by in situ hybridization and C-banding led to the conclusion that NR98116-9-2 was a 3R (3D) disomic substitution line and NR98116-9-3 was a 3R disomic addition line. The HMW-GS compositions were 1, 14 + 15, 6r + 8, 4 + 12 and 1, 14 + 15, 6r + 8, 5 + 10, respectively. It is very interesting that both lines may contain two Glu-B1 sites. The potential usefulness of the two germplasm that expressed seven different high molecular weight glutenin subunits in wheat quality improvement was discussed.

Chromosomes, Plant↗

[AFLP analysis on genetic diversity of wild Dactylis glomerata L. germplasm resources].

Thirty-seven accessions of wild Dactylis glomerata L. were used in assessment of genetic diversity by means of amplified fragment length polymorphism (AFLP). Nine primer pairs were selected from 64 preparations and then used to analyze the plant materials. A total of 400 DNA fragments were amplified, among which 336 (84.0%) were polymorphic. Genetic distance, from 0.0692-0.4214 among accessions, was calculated by the NTSYS-pc software. A dendrogram was constructed using UPGMA based on genetic distance matrix. Eight clusters were defined at the genetic similarity of 0.81. A two-dimensional PCO plot representing relationships among accessions was also produced by EIGEN, demonstrating the same distribution pattern as from the cluster analysis. Based on molecular analysis, the genetic variation of D. glomerata germplasms was closely associated with ploidy levels and geographical distributions.

Amplified Fragment Length Polymorphism Analysis↗

Research note: Efficient preservation of genome-edited chicken germplasm via N-methylacetamide-based semen cryopreservation.

The rapid development of genome-edited chicken lines for agricultural and biomedical use requires effective methods for long-term preservation. In birds, cryopreservation of oocytes and embryos is challenging due to the structure of the egg. This makes semen cryopreservation one of the most practical alternatives, however, this method has not been validated in genome-edited chickens. This study evaluated the feasibility of cryopreserving semen from genome-edited chickens using an N-methylacetamide (N-MA)-based protocol. Two genome-edited chicken lines were used in this study which are a DAZL-GFP germ cell reporter line and a RAG1 knockout line. Semen was stored for either one week or one month prior to assessing fertility and hatchability. The results demonstrated that cryopreserved semen from both genome-edited chicken lines maintained their fertilizing capacity, with no significant differences in fertility or hatchability compared to cryopreserved wild-type groups at either storage duration. Hatchability remained above 80% across all groups, indicating that semen cryopreservation did not significantly affect embryo development after successful fertilization. Furthermore, SYBR-14/PI staining revealed no significant differences in sperm viability between genome-edited and control groups, although both showed a significant decrease in viability compared to fresh semen. These findings demonstrate that N-MA-based semen cryopreservation provides a reliable and practical method for the preservation of genome-edited chicken lines.

Chicken↗

Virome of the Russian Grapevine Germplasm: A Final Study and Summary.

Ampelographic collections play an important role in the conservation of grapevine genetic resources and therefore require continuous phytosanitary monitoring. In this study, the virome of grapevines from the Magarach ampelographic collection in Russia was analyzed using total RNA high-throughput sequencing. A total of twenty-seven grapevine viruses and four viroids were identified. Two viruses were characterized as putative novel species: (+) ssRNA grapevine umbra-like virus 5 (GULV-5) and the bipartite (+) ssRNA grapevine Magarach secovirus (GMSV), which, together with related viruses, may represent a novel genus within the family Secoviridae. Among the economically important viruses, the most prevalent were grapevine fanleaf virus (76%), grapevine leafroll-associated virus 1 (39%), and grapevine virus A (33%). Mixed infections involving two or three of these viruses were detected in 50% of the analyzed grapevines. Grapevine virus D was detected in Russia for the first time. Phylogenetic analysis of 222 assembled virus and viroid genome sequences revealed high genetic diversity. The obtained results were summarized and compared with previous virome studies conducted on four Russian ampelographic collections.

RNA-Seq↗

Random amplified polymorphic DNA analysis of Indian scented basmati rice (Oryza sativa L.) germplasm for identification of variability and duplicate accessions, if any.

Genetic variation among the seventeen basmati rice accessions collected from the farmers' fields located at different places in India was analyzed using ten arbitrary tenmer oligonucleotide primers by the random amplified polymorphic DNA (RAPD) method. The number of polymorphic/monomorphic bands among the pairwise combinations of the accessions and the total number of bands were determined to categorize all the accessions in different groups of similarity and dissimilarity at the genetic level. The procedure described also allows the identification and discrimination of the individual genotypes of basmati rice including the identification of the duplicates in genetic resource collections.

DNA, Plant↗

Evaluation of Tolypothrix germplasm for phycobiliprotein content.

Twenty Tolypothrix strains, including 15 strains of T. tenuis, three strains of T. ceylonica and one strain each of T. nodosa and T. bouteillei, were evaluated for their phycobiliprotein content and composition. Significant differences among the Tolypothrix strains were found at both inter- and intra-specific levels in the production of phycobiliprotein constituents--phycocyanin (PC), allophycocyanin (APC) and phycoerythrin (PE). Four specific parameters, viz. PC or PE content, total phycobiliprotein and total protein content, and percentage of phycobiliproteins, in a mixture of total proteins were used to select four T. tenuis and one T. ceylonica strain as useful for phycobiliproteins production.

Bacterial Proteins↗

Genetic variation in the efficiency of nitrogen utilization and photosynthetic activity of flag leaves among the old and modern germplasm of winter wheat.

Genotypic variation in major components of the efficiency of nitrogen utilization and photosynthetic activity of flag leaves among old (released 1881-1963) and modern (released 1969-2003) cultivars of winter wheat was studied in field conditions under varied N fertilization levels (110, 90 and 80 kg N ha-1). Significant genotypic differences were observed for all characters. Their heritabilities ranged from 0.37 to 0.93 and were the lowest for the leaf efficiency of gas exchange, photosynthetic rate, straw N content and the economic index of N utilization efficiency (NUE). Some modern cultivars exhibited an enhanced tolerance to N shortage and several attributes of efficient N utilization (e.g. later senescing and more photosynthetically active flag leaves, increased ability to redistribute N into grains). The genotypes may serve as donors of appropriate characteristics for breeding. The observed cultivar-by-fertilization interactions suggest, however, that evaluations under diverse fertilization regimes may be necessary when searching for improved wheat efficiency and adaptation to less favourable environments.

Adaptation, Physiological↗

Genetic diversity of a germplasm collection of Cucurbita pepo using SRAP and AFLP markers.

Cucurbita pepo is a highly polymorphic species. The cultivars can be grouped into eight morphotypes in two subspecies, ssp. pepo and ssp. ovifera. A collection of 69 accessions representative of the morphotypes and some unclassified types was used for analysing the morphological and molecular diversity of this species. This collection includes commercial cultivars and Spanish landraces, which represent the great diversification of types that have arisen in Europe after this species arrived from America. For the molecular variability studies, two PCR-based systems were employed, AFLP and SRAP, which preferentially amplify ORFs. Principal coordinates analysis and cluster analysis using the UPGMA method clearly separate the accessions into the two subspecies through the use of both markers. However, the gene diversity and the genetic identity values among morphotypes and subspecies varied between the two marker systems. The information given by SRAP markers was more concordant to the morphological variability and to the evolutionary history of the morphotypes than that of AFLP markers. In ssp. ovifera, the accessions of the different morphotypes were basically grouped according to the fruit colour. This may indicate different times of development and also the extent of breeding in the accessions used. This study has allowed identification of new types that can be employed for the development of new cultivars. The landraces of the spp. ovifera, used as ornamental in Europe, have proved to be of great interest for preserving the diversity of C. pepo.

Cluster Analysis↗

Genetic diversity of oil palm (Elaeis guineensis Jacq.) germplasm collections from Africa: implications for improvement and conservation of genetic resources.

A total of 723 accessions of oil palm ( Elaeis guineensis Jacq.) from 26 populations representing ten countries in Africa and one Deli dura family were screened for allelic variation at seven enzyme loci from six enzyme systems using starch gel electrophoresis. On average, 54.5% of the loci were polymorphic (0.99 criterion). The average and effective number of alleles per locus was 1.80 and 1.35, respectively. Mean expected heterozygosity was 0.184, with values ranging from 0.109 (population 8, Senegal) to 0.261 (population 29, Cameroon). The genetic differentiation among populations was high (F(ST)=0.301), indicating high genetic divergence. The calculation of F(ST) by geographic zones revealed that the high F(ST) was largely due to F(ST) among populations in West Africa, suggesting diversifying selection in this region. The mean genetic distance across populations was 0.113. The lowest genetic distance (D) was observed between population 5 from Tanzania and population 7 from the Democratic Republic of the Congo (0.000) and the highest was found between population 4 from Madagascar and population 13 from Sierra Leone (0.568). The total gene flow across oil palm populations was low, with an Nm of 0.576, enhancing genetic structuring, as evident from the high F(ST) values. UPGMA cluster analysis revealed three main clusters; the western outlying populations from Senegal and Sierra Leone were in one cluster but separated into two distinct sub-clusters; the eastern outlying populations from Madagascar were in one cluster; the populations from Angola, Cameroon, The Democratic Republic of the Congo, Ghana, Tanzania, Nigeria and Guinea were in one cluster. The Deli dura family seems to be closely related to population 6 from Guinea. Oil palm populations with high genetic diversity-i.e. all of the populations from Nigeria, Cameroon and Sierra Leone, population 6 of Guinea, population 1 of Madagascar and population 2 of Senegal should be used in improvement programmes, whereas for conservation purposes, oil palm populations with high allelic diversity (A(e)), which include populations 22 and 29 from Cameroon, populations 39 and 45 from Nigeria, population 6 from Guinea, populations 5 and 13 from Sierra Leone and population 1 from Madagascar should be selected for capturing as much genetic variation as possible.

Africa↗

The development of lettuce backcross inbred lines (BILs) for exploitation of the Lactuca saligna (wild lettuce) germplasm.

Backcross inbred lines (BILs) were developed in which chromosome segments of Lactuca saligna (wild lettuce) were introgressed into L. sativa (lettuce). These lines were developed by four to five backcrosses and one generation of selfing. The first three generations of backcrossing were random. Marker-assisted selection began in the BC(4) generation and continued until the final set of BILs was reached. A set of 28 lines was selected that together contained 96% of the L. saligna genome. Of these lines, 20 had a single homozygous introgression (BILs), four had two homozygous introgressions (doubleBILs) and four lines had a heterozygous single introgression (preBILs). Segregation ratios in backcross generations were compared to distorted segregation ratios in an F(2) population, and the results indicated that most of the distorted segregations can be explained by genetic effects on pollen- or egg-cell fitness. By means of BIL association mapping we were able to map 12 morphological traits and hundreds of additional amplified fragment length polymorphic (AFLP) markers. The total AFLP map now comprises 757 markers. This set of BILs is very useful for future genetic studies.

Chromosome Mapping↗

AFLP reveals structural details of genetic diversity within cultivated olive germplasm from the Eastern Mediterranean.

Amplified fragment length polymorphism (AFLP) analysis was used to assess genetic inter-relationships among olive varieties cultivated in the Eastern Mediterranean Basin. The genotypes sampled included most of the important cultivars from Turkey, Greece and the Middle East and selected genotypes from the Western Mediterranean area. A total of 119 polymorphic markers were generated from five selective primer-pair combinations. The combined data sets generated by just two primer-pairs were adequate to discriminate between all 65 genotypes, while each primer-pair could individually identify up to 64 genotypes. A factorial correspondence analysis (FCA) plot indicated that the cultivars clustered into two relatively modestly defined groups. The first broad group was dominated by cultivars from Turkey but also included genotypes originating from the Middle East (Syria and Lebanon) that collectively formed a tight subcluster. The second group comprised Greek cultivars and those originating from the Western Mediterranean. A significant genetic distance value between Greek and Turkish cultivars was provided by an analysis of molecular variance (AMOVA). There was also evidence of substructure here, with an apparent separation of most Spanish and Italian clones. These findings are in general accordance to previous suggestions of an East-West divergence of olive cultivars, although the dichotomy is less extensive than reported previously and complicated by regional variation within each group.

Analysis of Variance↗

Rationalising germplasm collections: a case study for wheat.

In total 70 genebank accessions comprising 50 hexaploid, 12 tetraploid and 8 diploid wheats of the Gatersleben collection were selected based on the screening of the passport data for identical cultivar names or accession numbers of the donor genebanks. Twelve potential duplicate groups consisting of three to nine accessions with identical names/numbers were selected and analysed with DNA markers (microsatellites). A bootstrap approach based on re-sampling of both microsatellite markers and alleles within marker loci was used to test for homogeneity. Although several homogeneous groups were identified it became clear that cultivar name identity alone did not allow the determination of duplicates. A combination of SSR-analysis followed by the bootstrap method and database survey considering the botanical classification and other data (origin, growth habit and donor) available is recommended in order to determine duplicates. A procedure for the identification of duplicates and their further handling in ex situ genebanks is discussed.

Cluster Analysis↗

Generation and characterization of low phytic acid germplasm in rice (Oryza sativa L.).

Phytic acid (PA, myo-inositol 1,2,3,4,5,6-hexakisphosphate), or its salt form, phytate, is commonly regarded as the major anti-nutritional component in cereal and legume grains. Breeding of low phytic acid (lpa) crops has recently been considered as a potential way to increase nutritional quality of crop products. In this study, eight independent lpa rice mutant lines from both indica and japonica subspecies were developed through physical and chemical mutagenesis. Among them, five are non-lethal while the other three are homozygous lethal. None of the lethal lines could produce homozygous lpa plants through seed germination and growth under field conditions, but two of them could be rescued through in vitro culture of mature embryos. The non-lethal lpa mutants had lower PA content ranging from 34 to 64% that of their corresponding parent and four of them had an unchanged total P level. All the lpa mutations were inherited in a single recessive gene model and at least four lpa mutations were identified mutually non-allelic, while the other two remain to be verified. One mutation was mapped on chromosome 2 between microsatellite locus RM3542 and RM482, falling in the same region as the previously mapped lpa1-1 locus did; another lpa mutation was mapped on chromosome 3, tightly linked to RM3199 with a genetic distance of 1.198 cM. The latter mutation was very likely to have happened to the LOC_Os03g52760, a homolog of the maize myo-inositol kinase (EC 2.7.1.64) gene. The present work greatly expands the number of loci that could influence the biosynthesis of PA in rice, making rice an excellent model system for research in this area.

Alleles↗

Foliar disease resistance phenomics of fungal pathogens: image-based approaches for mapping quantitative resistance in cereal germplasm.

Host plant resistance is the most effective and environmentally sustainable means of reducing yield losses caused by fungal foliar pathogens of cereal species. Cereal genebank collections hold diverse pools of potentially underutilized disease resistance alleles, and cereal genomic resources are well advanced due to large-scale sequencing and genotyping efforts. Genome-Wide Association Studies (GWAS) have emerged as the predominant association genetics technique to initially discover novel disease resistance loci or alleles in these diverse collections. Traditional disease resistance phenotyping methods are reliant on visual estimation of disease symptom severity and have successfully supported genetic mapping studies either via GWAS or QTL mapping in biparental populations facilitating both marker development and gene cloning efforts. Due to foliar pathogens having a high capacity to evolve, there is a need to pyramid disease resistance genes with diverse mechanisms for durable control. Resistance expressed as a quantitative trait, known as quantitative resistance (QR), is hypothesized to be more durable, unlike major R-gene resistance that is race-specific and can be vulnerable to breaking down without gene stewardship. However, assessing QR visually is challenging, particularly when complicated by complex genotype × environment (G × E) effects in the field. High-throughput image-based phenotyping provides accurate and unbiased data that can support foliar disease resistance screening efforts of genebank collections using GWAS. In this review, we discuss image-based disease phenotyping based on macroscopic (visible symptoms) and microscopic features during the host-pathogen interaction. Quantitative image analysis approaches using conventional and artificial intelligence (AI) algorithms are also discussed.

Disease Resistance↗

Allelic variation at (TAA)n microsatellite loci in a world collection of chickpea (Cicer arietinum L.) germplasm.

A set of 12 randomly selected (TAA)n microsatellite loci of the cultivated chickpea (Cicer arietinum L.) were screened in a worldwide sample comprising 72 landraces, four improved cultivars and two wild species of the primary gene pool (C. reticulatum and C. echinosperum) to determine the level and pattern of polymorphism in these populations. A single fragment was amplified from all the accessions with each of 12 sequence-tagged microsatellite site markers, except for one locus where no fragment was obtained from either of the two wild species. There was a high degree of intraspecific polymorphism at these microsatellite loci, although isozymes, conventional RFLPs and RAPDs show very little or no polymorphism. Overall, the repeat number at a locus (excluding null alleles) ranged from 7 to 42. The average number of alleles per locus was 14.1 and the average genetic diversity was 0.86. Based on the estimates obtained, 11 out of the 12 frequency distributions of alleles at the loci tested can be considered to be non-normal. A significant positive correlation between the average number of repeats (size of the locus) and the amount of variation was observed, indicating that replication slippage may be the molecular mechanism involved in generation of variability at the loci. A comparison between the infinite allele and stepwise mutation models revealed that for 11 out of the 12 loci the number of alleles observed fell in between the values predicted by the two models. Phylogenetic analysis of microsatellite polymorphism in C. arietinum showed no relationship between accession and geographic origin, which is compatible with the recent expansion of this crop throughout the world.

Alleles↗