Chloroplast DNA molecular characterization and leaf volatiles analysis of mint (Mentha; Lamiaceae) populations in China

Non-coding chloroplast DNA (cpDNA) was used to evaluate genetic diversity and relationships within the section Mentha collected from China. Leaf volatiles of Mentha spicata accessions and its parent were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). To this end, genetic diversity of 30 Mentha accessions from different geographic origins of China, representing six species and two hybrids, was assessed. Molecular studies grouped the samples mainly according to genetic relationship established by conventional methods. Cluster analysis based on the leaf volatiles chemical composition of M. spicata accessions defined two main chemotypes: 1,8-cineole-piperitenone oxide type and limonene-carvone type. The results suggested that there was a high genetic variability among individuals of Mentha in China. A distinct correlation between cpDNA marker and volatile oils in M. spicata accessions was revealed. Moreover, the chemical composition followed maternal inheritance in M. spicata accessions. In conclusion, the genetic diversity of Mentha populations as shown in this study should play a critical role in future selection and breeding programs.     

甘蓝型油菜微核心种质耐旱鉴定与评价指标筛选

干旱是威胁油菜生产的重要非生物逆境，建立有效的耐旱鉴定方法，筛选鉴定耐旱油菜种质资源，对培育油菜耐旱新品种、提高油菜稳产性具有重要意义。本文以107份甘蓝型油菜微核心种质为材料，设置正常供水和干旱2个处理，采用干旱-复水-干旱-复水的处理方法，通过测定旱害指数、叶片萎蔫指数、地上部鲜重胁迫指数、地上部干重胁迫指数、根鲜重胁迫指数、根干重胁迫指数、植株总鲜重胁迫指数、植株总干重胁迫指数等指标，评价油菜种质耐旱性变异，筛选理想的耐旱评价指标。结果表明，干旱严重抑制了甘蓝型油菜的生长，8个评价指标均表现出显著差异，并呈正态分布。地上部鲜重胁迫指数、植株总鲜重胁迫指数和旱害指数3个指标之间显著相关，可作为甘蓝型油菜苗期耐旱性的主要评价指标，其中地上部鲜重胁迫指数最为简便、准确和有效；综合应用这3个评价指标，明确了107份微核心种质耐旱性，筛选出2份高耐旱种质，为耐旱相关研究及育种提供了评价方法和材料。     

Genetic diversity of accessions of Iranian Aloe vera based on horticultural traits and RAPD markers

In this study, genetic diversity of 10 accessions of Aloe vera, collected from different parts of Iran, were evaluated using horticultural and random amplified polymorphic DNA (RAPD) data. Statistical analysis showed significant differences for all horticultural characteristics among the accessions, suggesting that selection for relevant characteristics could be possible. The analysis of molecular diversity was used the RAPD technique, with 10 random primers of 10-mer oligonucleotides. Out of the 10 primers, five were polymorphic, producing 269 DNA bands, 189 of which were polymorphic among accessions. A dendrogram was prepared on the basis of a similarity matrix using the Unweighted Pair Group Method Arithmetic Mean (UPGMA) algorithm, separating the 10 accessions into two groups. Results show that both environmental and genetic factors are effective in observing variations. Our results also indicate that the RAPD approach, along with horticultural analysis, seemed to be best-suited for assessing with high accuracy the genetic relationships among distinct A. vera accessions.     

Multiple-disease resistance in Vicia faba: Multi-environment field testing for identification of combined resistance to rust and chocolate spot

The aim of this work was to identify Vicia faba germplasm resistant both to rust and chocolate spot. A collection of 43 accessions of V. faba previously identified as chocolate-spot resistant was evaluated for rust and chocolate spot resistance in Egypt and in Spain. The genotype and genotype × environment (GGE) biplot analyses allowed the selection of 11 accessions resistant to both diseases in those locations. These 11 accessions were evaluated for rust in an additional field trial in Spain, all performing better than the susceptible check. The joint analysis of the 11 accessions in the four field trials where they had been evaluated revealed no significant effects either for genotype, environment or the genotype × environment interaction. They are hence promising sources of resistance, both for their low severities and their stable responses across the studied environments. This collection was also tested under controlled conditions against the Egyptian and the Spanish isolates of rust present in the field trials further underlining the stable performance of these genotypes. Finally, the effect of previous infection with chocolate spot on rust resistance was assessed under controlled conditions and no influence of it was found.     

Genetic Diversity Analysis of Rice Germplasm in Tripura State of Northeast India Using Drought and Blast Linked Markers

We genotyped 74 rice germplasms including Tripura's local landraces, improved varieties, cultivars and breeding lines and other rice varieties using molecular markers for genetic diversity, drought QTLs, and blast resistance genes. The number of alleles per locus ranged from 2 to 5 with an average of 2.9. The polymorphic information content value per locus ranged from 0.059 (RM537) to 0.755 (RM252) with an average of 0.475. Cluster analysis based on 30 simple sequence repeat markers revealed 5 clusters and also indicated the presence of variability within the rice accessions. The drought QTL qDTY2.1 was found in 56.0% of germplasms and qDTY1.1 was detected in only 6.8% of the germplasms. Out of seven rice blast resistance genes screened, only two rice varieties, RCPL-1-82 and Buh Vubuk (Lubuk), were positive for four blast resistance genes while only Releng possessed two blast resistance genes. Among 74 rice germplasms, only three accessions, Releng, RCPL1-82 and Buh Vubuk (Lubuk), possessed both drought-related QTLs and blast resistance genes. Overall, the 74 indigenous rice genotypes showed low level of genetic diversity, which is in contrast to high level of genetic diversity among rice varieties in northeast India, where highlights the good farming practice, conservation of germplasms and the limitation of molecular markers employed in this study. The presence of both drought related QTLs and blast resistance genes in some of the germplasms can be useful in future breeding programmes.     

Breeding prospects of Lunaria annua L.

Lunaria annua is a biennial cruciferous oil seed crop. The seeds contain 30–35% oil, which consists of 67% long chain fatty acids (44% erucic acid, C22:1, and 23% nervonic acid, C24:1). The oil is suitable as lubricant. In addition, recent developments indicate that nervonic acid may be used as raw material for the production of a medicine against multiple sclerosis. The biennial character of Lunaria is a main constraint for an economically feasible production of Lunaria oil. The crop has to be sown early in the summer to achieve vigorous plant development required for vernalization during the winter. It would be a great advance when Lunaria could be sown later in the summer after an early harvested crop. From 1993, breeding research in Lunaria has been performed at CPRO-DLO, Wageningen, The Netherlands. A collection of 76 accessions was maintained and evaluated for agronomic performance. In 1995, a selection of 24 accessions were sown at four sowing dates, from end June until end of August and in 1996, 12 promising accessions were sown again at 15 and 30 July. Three accessions showed suitability for delayed sowing until beginning of August. Delay in sowing time caused also delay of flowering and seed ripening. Seed yield amounted to 1200 kg/ha in 1996 and 1700 kg/ha in 1997. Seed oil content varied from 30–38%. Severe infection of Albugo candida and Alternaria occurred and might have reduced seed yield in both years. Most accessions sustained severe winter frost of−17°C very well. It is concluded that within the evaluated gene pool promising variation is available to select for ability for late sowing, to improve the disease resistance and to increase seed yield and seed oil content.     

Molecular relationships between Australian annual wild rice,Oryza meridionalis,and two related perennial forms

Background

The perennial, Oryza rufipogon distributed from Asia to Australia and the annual O. meridionalis indigenous to Australia are AA genome species in the Oryza. However, recent research has demonstrated that the Australian AA genome perennial populations have maternal genomes more closely related to those of O. meridionalis than to those found in Asian populations of O. rufipogon suggesting that the Australian perennials may represent a new distinct gene pool for rice.

Results

Analysis of an Oryza core collection covering AA genome species from Asia to Oceania revealed that some Oceania perennials had organellar genomes closely related to that of O meridionalis (meridionalis-type). O. rufipogon accessions from New Guinea carried either the meridionalis-type or rufirpogon-type (like O. rufipogon) organellar genomes. Australian perennials carried only the meridionalis-type organellar genomes when accompanied by the rufipogon-type nuclear genome. New accessions were collected to better characterize the Australian perennials, and their life histories (annual or perennial) were confirmed by field observations. All of the material collected carried only meridionalis-type organellar genomes. However, there were two distinct perennial groups. One of them carried an rufipogon-type nuclear genome similar to the Australian O. rufipogon in the core collection and the other carried an meridionalis-type nuclear genome not represented in the existing collection. Morphologically the rufipogon-type shared similarity with Asian O. rufipogon. The meridionalis-type showed some similarities to O. meridionalis such as the short anthers usually characteristic of annual populations. However, the meridionalis-type perennial was readily distinguished from O. meridionalis by the presence of a larger lemma and higher number of spikelets.

Conclusion

Analysis of current accessions clearly indicated that there are two distinct types of Australian perennials. Both of them differed genetically from Asian O. rufipogon. One lineage is closely related to O. meridionalis and another to Asian O. rufipogon. The first was presumed to have evolved by divergence from O. meridionalis becoming differentiated as a perennial species in Australia indicating that it represents a new gene pool. The second, apparently derived from Asian O. rufipogon, possibly arrived in Australia later.     

Genome-wide association study identifies novel loci and candidate genes for drought stress tolerance in rapeseed

Rapeseed(Brassica napus)is one of the most important oil crops worldwide;however,drought seriously curtails its growth and productivity.Identifying drought-tolerant germplasm is an efficient strategy for addressing water shortages.Here,we phenotyped a panel of 264 B.napus accessions at full-bloom stage using water loss ratio(WLR)as drought-tolerant index.It identified 8 low-WLR and 6 high-WLR accessions,regarded as droughttolerant and drought-sensitive,respectively.Comparing with drought-sensitive accessions at the seedling stage,drought-tolerant accessions had shown better performance in maintaining fresh and dry weights,and performed the higher expression of drought-induced marker genes under drought stress.Subsequently,a total of 139 SNPs(single nucleotide polymorphisms)were identified associated with the WLR using a genome-wide association study(GWAS)among 264 B.napus accessions,with the largest number SNPs at chromosome A10,and 13 SNPs significantly were associated with the WLR(-log₁₀(p-value)>6).Furthermore,4 putative candidate genes(BnaC09.RPS6,BnaC09.MATE,BnaA10.PPD5 and BnaC09.Histone)were screened involving in drought tolerance in B.napus.Together,our results highlight the WLR's importance in drought tolerance and establish the foundation for improving WLR-associated drought tolerance in rapeseed.     

Spatial analysis of genetic diversity in a comprehensive collection of the native grass Bromus auleticus Trinius (ex Nees) in Uruguay

Bromus auleticus Trinius (ex Nees) is a perennial allogamous C3 grass native to the Campos grasslands. It has potential to play a key role in restoration of both abandoned agricultural and overgrazed lands. We aim to reveal the existence of local adaptation and to assess whether phenotypes could be differentiated in relation to their ecogeographic region of origin. Previous reports estimated significant levels of variability among and within populations, but as yet there is no evidence of local adaptation and phenotypic relations with ecogeographic regions for this species. Indeed, there is a lack of knowledge about the spatial distribution of diversity, which limits the possibilities of using B. auleticus in ecological restoration and plant breeding. A comprehensive collection of 82 accessions of B. auleticus from eight ecogeographic regions within Uruguay was used to characterize diversity by measuring phenologic, morphologic, productive and reproductive traits. We found high diversity among B. auleticus accessions, and the spatial distribution of this diversity follows a pattern accounted by ecogeographic regions to a large extent. Our results also indicate large genotype × environment interaction, confirming local adaptation. Phenotypic values permitted, with a 3.7% error rate, discrimination of accessions according to their ecogeographic region of origin, suggesting the existence of differentiated ecotypes. We consider that the diversity observed should be conserved and used for breeding cultivars of this species and for the restoration of degraded grasslands by considering ecogeographic regions when selecting seed sources.     

Molecular analyses of genetic variability in soap nut (Sapindus mukorossi Gaertn.)

Sapindus mukorossi Gaertn. (Sapindaceae) is commercially an important plant much valued for its fruits commonly known as soap nut as well as for the seed oil used as a feed-stock for biodiesel production. Molecular polymorphism, determined, using three single primer amplification reaction (SPAR) methods (RAPD, DAMD and ISSR) is used to assess the genetic variability in 69 accessions of S. mukorossi Gaertn., encompassing different geographical regions in India. Cumulative band data generated using the three SPAR methods resulted in 82.49% polymorphism across all genotypes of S. mukorossi Gaertn. UPGMA tree showed two major clusters that were in accordance with the geographical diversity. The intra-population genetic diversity (0.16), Shannon information index (0.24) and polymorphic loci (45.62%) were highest in AS population, while the corresponding lowest values were recorded in HP population. The inter-population average distance ranged from 0.05 (GA and HP) to 0.26 (AP and HP). Based on these distances, the UPGMA tree for the populations was computed, which showed the presence of two major clusters. Cluster I contained all the populations from Himachal Pradesh (HP) and Uttarakhand (GA and KU) and cluster II grouped the populations from North Eastern region (AS, AP, MG). The present study showed that SPAR methods are informative and useful to unravel the diversity among different populations of the soap nut plants and is a prelude for further utilization of promising and genetically divergent materials in the breeding programmes.     

Genetic characterization of kernel polyphenol oxidases in wheat and related species

Polyphenol oxidase (PPO) activity causes undesirable darkening of raw Asian noodles and other wheat products. In this study we investigate the genetic origins and diversity of wheat kernel PPO. PPO was characterized via activity assays, antigenic staining, and Southern blots in Triticum aestivum, Triticum dicoccoides, Triticum durum, Triticum dicoccum, Triticum monococcum, Triticum urartu, Aegilops speltoides, and Aegilops tauschii. Among these species, PPO activity was well-correlated with antigenic staining intensity toward a wheat kernel-type PPO antibody. High PPO activity was observed in all three T. monococcum accessions (A^m genome), one Ae. speltoides accession, one T. durum accession, and two hexaploid wheat cultivars. Southern blots suggested the presence of two or more kernel-type PPO genes in diploid progenitors of the hexaploid A, B, and D genomes. Whole-kernel PPO activity was evaluated in disomic substitution lines derived from three T. dicoccoides accessions in the background of T. durum ‘Langdon’. PPO activity was primarily associated with chromosome 2A and to a much lower degree with chromosome 2B. DNA sequence comparisons showed that the intron associated with the high PPO allele on chromosome 2AL of hexaploid wheat had 94% nucleotide identity with the homeologous intron found in T. monococcum, a species with high kernel PPO activity. This implies that the ancestral PPO allele on the A genome is one of the high activity, and the low PPO allele found in hexaploid wheat represents a relatively recent genetic alteration. Results confirm the presence of multiple kernel-type PPO genes in the diploid and tetraploid progenitors and relatives of hexaploid wheat. However, it is likely that relatively few of the many kernel-type PPO genes present in wheat contribute substantially to kernel PPO activity. A single genetic locus on homeologous group 2 chromosomes may be the primary cause of high PPO activity in wheat kernels.     

Manihot flabellifolia Pohl,wild source of resistance to the whitefly Aleurotrachelus socialis Bondar (Hemiptera: Aleyrodidae)

Aleurotrachelus socialis is one of the most important pests of cassava (Manihot esculenta Crantz) in the Neotropics. In Colombia, high whitefly populations can reduce crop yields by 79%; and although the farmers intensify the use of insecticides, this practice is highly contaminating, costly and leads to the development of resistance in the insect. An alternative for managing whitefly populations is to develop genetically resistant varieties. Wild parents of Manihot are a useful source of genes against pests for the cultivated species of cassava. Based on prior research that showed the existence of moderate-to-high levels of resistance to A. socialis in Manihot flabellifolia, a wild species of cassava, this study was proposed to characterize this new source of resistance, evaluating the biology and demographics of A. socialis on eight accessions of M. flabellifolia, a susceptible check (CMC-40) and a resistant (MEcu72) check. The averages of A. socialis longevity and fecundity on the accessions were not significantly different to MEcu72, but different from CMC-40 (P < 0.05). Development time was not significantly different, ranging from 35–40 days on accessions and MEcu72 and 33.5 days on CMC-40 (P < 0.05). In contrast, the population growth rate (r_m) was significantly lower on the M. flabellifolia accessions, with Fla 61 standing out with a growth rate 98 and 99% less than that obtained on MEcu72 and CMC-40, respectively. Once the resistant levels have been identified to A. socialis on the M. flabellifolia accessions, interspecific crosses of M. esculenta subsp. M. flabellifolia and backcross programs could be developed to incorporate the desirable characteristics from the wild relatives into elite progenitors of M. esculenta.     

Genetic and morphological characterization of Acroceras macrum Stapf

Acroceras macrum is a warm‐season grass used for pastures, and it has high nutritive value. The scarce genetic and reproductive information about this grass is limiting its breeding for forage. The objective of this research was to determine the number of genotypes and cytotypes present in a collection established in Argentina, and to estimate the diversity of this species based on molecular markers, and morphological and agronomic characteristics. The number of genotypes among forty‐seven accessions was determined using ISSR markers. The identified genotypes were further characterized for ploidy levels. In addition, the genetic distances among them were estimated with ISSR markers. They were also propagated and planted into the field following a randomized complete block design with four replications, and a group of sixteen morphological and agronomic variables were evaluated. A total of twenty‐seven genotypes were identified in this collection; twenty‐two genotypes were tetraploid (2n = 4x = 36) and five were hexaploid (2n = 6x = 54). These two cytotypes were found clearly separated by both genetic and phenotypic characteristics. Above‐ground biomass, internode length, initial growth and number of spikelets per inflorescence were the most variable traits. The high diversity present in this collection will allow for the initiation of a breeding programme for A. macrum.     

Fire blight (Erwinia amylovora) resistant/susceptibility of native apple germplasm from eastern Turkey

This study aimed to determine the resistance/susceptibility to fire blight of apple germplasm resources from the province of Erzincan in eastern Turkey. In total, 32 native apple accessions from four regions within the province were tested under greenhouse conditions by inoculating the shoot tips with pathogenic bacterium. Shoot tips were wounded for inoculation. Genotypical susceptibility index (GSI%) values were computed for each genotype based on the length of the lesion that developed on each shoot. Accessions were grouped into five classes of resistance/susceptibility, as follows: Class A (resistant); Class B (moderately resistant); Class C (moderately susceptible); Class D (susceptible); and Class E (highly susceptible). GSI% values differed significantly among accessions (p < 0.01). Five accessions received a rating of Class A, and 7 accessions received a rating of Class B. This was followed by 9 accessions with a rating of Class C, 5 accessions with a rating of Class D and 6 accessions with a rating of Class E. The findings of this study are expected to contribute to breeding efforts with respect to apple resistance to fire blight.     

Resistance to Penicillium allii in accessions from a National Plant Germplasm System Allium collection

Multiple Allium accessions (garlic, and wild and ornamental Allium species) were screened for resistance using Penicillium allii and A. sativum (positive control). Single accessions of A. aflatunense, A. atroviolaceum, A. stipitatum, and Allium sp. remained asymptomatic. Single accessions of A. roseum and A. senescens, two accessions each of A. acuminatum and A. ampeloprasum and a single accession of A. moly displayed lesion expansion rates not exceeding 22%, 26%, 46%, 50%, 61%, 67% and 67%, respectively, of positive controls. Single accessions of A. sativum var. ophioscordon and A. scorodoprasum displayed rates not exceeding 68% and 55%, respectively, of positive controls with deep wounding, but did not consistently differ with shallow wounding. Accessions of A. canadense, A. sativum or A. longicuspis did not differ, differed inconsistently, or differed insubstantially from positive controls. Lesion expansion rates for A. acuminatum, A. ponticum and A. scorodoprasum were significantly less than in positive controls, but their small bulbs often rotted completely. Results document publicly available germplasm possessing significant resistance to P. allii.     

Genetic Diversity and Redundancy Among Potato Accessions in the Montenegrin Collection as Revealed by Microsatellite Markers

Potato was introduced in Montenegro in the middle of the eighteenth century. Since then it has become the most important crop in plant production. During the period between 2008 and 2010 a total of 52 potato accessions was collected across Montenegro and stored in a national gene bank. In the study reported here 23 accessions from the collection were examined using microsatellite (also known as simple sequence repeats or SSRs) molecular markers with the aim to explore genetic diversity and redundancy within the germplasm. The accessions were selected on the basis of preliminary characterization of all 52 accessions for 11 lightsprout traits. Molecular characterization of 23 accessions by 12 SSR markers was carried out at SASA (Science and Advice for Scottish Agriculture) that manages a database of more than 3000 genetic profiles of potato from Europe and abroad. Comparison of SSR genetic profiles of Montenegrin collection against the existing SASA database allowed us to test the authenticity of the Montenegrin accessions. Out of the 23 accessions examined, 13 showed distinct genetic profiles of which seven showed perfect matching with known cultivars, two profiles showed strong similarity to another two cultivars, and four profiles were found unique with regards to the SASA database. Application of microsatellite markers in this study provided valuable information on the extent of genetic diversity residing within Montenegrin potato germplasm; it gave clear indications of the scale of redundancy within the collection; and helped clarify the identity of the accessions. Four accessions within the collection might incorporate unique variation and will be subjected to further agronomical examinations to assess their potential for breeding purposes.     

Discriminant analysis of selected yield components and fatty acid composition of chosen Triticum monococcum, Triticum dicoccum and Triticum spelta accessions

This study analyses the variability of key yield components, the content of protein and crude fat in grain and the fatty acid composition of 50 spring accessions of Triticum monococcum, Triticum dicoccum and Triticum spelta of various origins. The average protein content of the grain of T. monococcum was 20.8%, of T. dicoccum 19.7%, and of spelt 17.0%. The crude fat content of T. monococcum grain (2.7%) was significantly higher compared with T. spelta (2.4%) and T. dicoccum (2.3%). In crude fat, fatty acids C18:2, C18:1 and C16 predominated. T. spelta was characterised by the highest concentrations of C18:2 and C16 (55.89% and 18.77% respectively), while T. monococcum had the highest content of C18:1 (26.35%). The structure of analysed fatty acids proved to be highly desirable in this species. A discriminant analysis performed separately for five variables: protein and fat content and three biometrical characters and separately for fatty acid composition enabled three Triticum species to be distinguished. These species also differed significantly with respect to the C18:1/C16 ratio which was equal to 1.78, 1.06, 1.47 and 0.99 in T. monococcum, T. dicoccum, T. spelta and Triticum aestivum respectively.     

Sponge-Derived Kocuria and Micrococcus spp. as Sources of the New Thiazolyl Peptide Antibiotic Kocurin

Forty four marine actinomycetes of the family Microccocaceae isolated from sponges collected primarily in Florida Keys (USA) were selected from our strain collection to be studied as new sources for the production of bioactive natural products. A 16S rRNA gene based phylogenetic analysis showed that the strains are members of the genera Kocuria and Micrococcus. To assess their biosynthetic potential, the strains were PCR screened for the presence of secondary metabolite genes encoding nonribosomal synthetase (NRPS) and polyketide synthases (PKS). A small extract collection of 528 crude extracts generated from nutritional microfermentation arrays was tested for the production of bioactive secondary metabolites against clinically relevant strains (Bacillus subtilis, methicillin-resistant Staphylococcus aureus (MRSA), Acinetobacter baumannii and Candida albicans). Three independent isolates were shown to produce a new anti-MRSA bioactive compound that was identified as kocurin, a new member of the thiazolyl peptide family of antibiotics emphasizing the role of this family as a prolific resource for novel drugs.