A phenetic analysis of morphological variation in Citrullus lanatus in Namibia

Morphological data recorded from field trials using Citrullus lanatus germplasm collected in Namibia were used to analyse and compare the various morphotypes of this species. The experiment comprised wild types and local landraces as well as commercial cultivars. Cluster analysis supported the indigenous classification system used in Namibia, in which Citrullus types are distinguished based on gross morphology, ecology and usage and grouped into seed, cooking and fresh-eating (watermelon) types. Commercial watermelon cultivars formed a distinct cluster. Wide variation was found within the local types whereas the genetic basis of the commercial type appears to be narrow. The commercial cultivars were most closely related to local watermelon types and more distantly related to the wild types, whereas the cooking melons form an intermediate group.     

Genetic diversity of cultivated flax (Linum usitatissimum L.) and its wild progenitor pale flax (Linum bienne Mill.) as revealed by ISSR markers

Little is known about the genetic diversity of pale flax (Linum bienne Mill.), the wild progenitor of cultivated flax (L. usitatissimum L.), and ex situ germplasm of pale flax was scarce. Effort was made to collect 34 pale flax accessions and five landrace accessions of cultivated flax in Turkey. The inter simple sequence repeat (ISSR) technique was applied to characterize this set of flax germplasm, along with one Turkish cultivar, one Russian cultivar, five winter and four dehiscent type accessions of cultivated flax. Twenty-four ISSR primer pairs detected a total of 311 DNA fragments, of which 298 bands were polymorphic across 493 flax samples (roughly 10 samples per accession). These polymorphic bands had frequencies ranging from 0.002 to 0.998 and averaging 0.38. Accession-specific ISSR variation (Fst values) ranged from 0.469 to 0.514 and averaged 0.493. There was 49.3% ISSR variation resided among these 50 accessions, 35.9% harbored among landrace, winter, dehiscent types of cultivated flax and pale flax, and 38.2% present among 34 pale flax accessions. Pale flax displayed more ISSR variation than landraces and dehiscent type, but less than winter type, of cultivated flax. Clustering 493 individual plants revealed that these assayed plants were largely grouped according to their plant types and that pale flax was genetically more close to the dehiscent type, followed by the winter type and landrace, of cultivated flax. Pale flax collected within the geographic range of 180 km displayed a significant spatial genetic autocorrelation. Genetic distances among the pale flax accessions were significantly associated with their geographic distances and elevation differences. These findings are significant for understanding flax domestication and its primary gene pool.     

Target region amplification polymorphism (TRAP) for assessing genetic diversity and marker-trait associations in chickpea (Cicer arietinum L.) germplasm

The effective utilization of crop diversity held in genebanks depends on our knowledge of useful traits and available markers associated with the target traits. Target region amplification polymorphism (TRAP) was used to evaluate the genetic diversity and underlying relationships among 263 accessions of chickpea landraces maintained by the USDA-ARS Western Regional Plant Introduction Station in Pullman, WA, USA. Two-hundred sixty-two TRAP markers were amplified by eight primer combinations. Altogether, 110 (42 %) markers were polymorphic, the other 152 (58 %) displayed no variation. These polymorphic markers revealed important differences among the accessions, with an estimated, mean pair-wise genetic distance of 25.82 %, ranging from 2.8 to 50.0 %. Genetic distance analysis divided the accessions into two major groups, with 113 and 150 accessions each, and substantial association between molecular diversity and geographic origin was evident. Bayesian analysis of population structure revealed two groups (K = 2) with evidence for six sub-groups. Additionally, the population structure of a subset of 110 lines was determined (K = 3) for testing marker-trait associations (MTAs). Phenotypic traits included the concentrations of protein and nine mineral elements in the seeds. Two MTAs were significant (p < 0.01) for concentrations of Ca and K, and three MTAs were significant for Cu and Ni concentrations. The results indicate that this population is useful for genome-wide association studies on other economic traits given the level of genetic diversity uncovered and the marker-trait associations in seed minerals discovered.     

Complexity and adaptability of a traditional agricultural system: case study of a gall midge resistant rice landrace from northern Thailand

Adaptability of traditional agricultural systems is suggested by their success over time, but documentation of how this happens is rare. This paper shows how genetic diversity in a rice landrace enables rice farming system of northern Thailand to adapt to a constraint of an insect pest, microenvironments of mountainous landscape and people’s different tastes in rice. Resistance to laboratory-reared gall midge varied among accessions the rice landrace Muey Nawng and gall midge populations. Higher rice yield in farmers’ fields reflected adaptation to local environment as well as resistance to gall midge. Microsatellite variation of the accessions correlated negatively with their gall midge resistance, but there was also variation in heading time and endosperm starch. Presence of non-waxy endosperm in glutinous rice provides opportunity to select for rice that is cooked into non-glutinous rice preferred by minority groups who live at higher elevations, where the gall midge is emerging as a new threat, possibly because of climate change. These data show how genetic diversity of a rice landrace coupled with seed management by farmers enabled a rice farming system to adapt to the varied microenvironment of a mountainous landscape under the constraint of an insect pest and people’s different tastes in rice.     

Differentiation between fiber and drug types of hemp (Cannabis sativa L.) from a collection of wild and domesticated accessions

Accessions of wild and domesticated hemp (Cannabis sativa L.) originating from Colombia, Mexico, California, Bolivia, Thailand, Afghanistan, Serbia, Hungary, south Africa and different regions of China, were studied by means of DNA polymorphisms in order to discriminate between drug and fiber types. Analysis of molecular variance (AMOVA) was used to partition the total genetic variance within and among populations. The significance of the variance components was tested by calculating their probabilities based on 999 random permutations. AMOVA revealed 74 % variation among accessions and 26 % within accessions, all AMOVA variation was highly significant (P < 0.001). The cluster analysis of molecular data, grouped accessions into eight clusters and gave a matrix correlation value of r = 0.943, indicating a very good fit between the similarity values implied by the phenogram and those of the original similarity matrix. In this study, DNA polymorphisms could discriminate the fiber and drug types, and accessions were grouped in accordance to their classification and uses. In addition, seed size variation and micromorphological characters of seeds were studied by means of a scanning electron microscope (SEM). Seeds varied significantly in size, and were bigger in the fiber types. SEM analysis exhibited variation of micromorphological characters of seeds that could be important for discriminating the fiber or drug types.     

The Pacific yam (<Emphasis Type="Italic">Dioscorea nummularia</Emphasis> Lam.), an under-exploited tuber crop from Melanesia

This study aims to explore the potential of Dioscorea nummularia Lam. and to gather information as a prelude to its improvement by plant breeding. After a taxonomic clarification and a review of the botanical data, its geographical areas of origin and distribution are determined. A germplasm survey conducted on nine islands of Vanuatu enabled the collection of 110 accessions corresponding to 84 cultivars. These were grouped into: (1) wild forms under domestication, (2) annual types, (3) perennial-type cultivars of D. nummularia and (4) natural interspecific hybrids with D. alata L. Within each group, remarkable morphological variation exists for stem spininess, leaf shape, emerging leaf colour, tuber shape and tuber flesh colour. Selected accessions examined for ploidy had chromosome numbers ranging from 2n = 3x = 60 to 2n = 6x = 120. Chloroplast counts conducted on stomatal guard cells confirmed the variation in ploidy levels. Eight different male cultivars were studied for pollen fertility, but <5% of the pollen grains appeared to be fertile when stained with acetocarmine. However, profuse flowering and synchrony favour pollination and result in fruit set. All accessions are resistant to anthracnose. Viruses were detected but are morphologically not discernible, with no incidence on yield. Physicochemical analyses of tubers from 16 accessions revealed that D. nummularia and interspecific hybrids have high percentages of dry matter (mean of 33.11%) and starch (82.81%). The Pacific yam could be improved through conventional hybridisation or used for interspecific crosses with related economically important species. Recommendations are made for further research.     

A study of variability in the Sicilian faba bean landrace ‘Larga di Leonforte’

In this paper molecular analysis, physical, technological, and chemical traits were used to estimate the level of variation on five accessions of a locally adapted Sicilian faba bean (Vicia faba L.) landrace named ‘Larga di Leonforte’. DNA analysis was performed using the Amplified Fragment Length Polymorphism (AFLP) molecular marker class and two other faba beans (cv. ‘Aguadulce Samba’—Spain—and landrace ‘Locale di Modica’—Sicily) were used as controls. Although the accessions of ‘Larga di Leonforte’ varied significantly for most of the agronomical and physical traits, this landrace generally had a heavy seed weight, short but large pods, and no more than two seeds per pod. This last characteristic allowed for erect pod angle attitude at maturity. Idratation data showed difference among accessions in seed weight at full hydration and in absorption rate at the very beginning of the hydration process, while any difference among accessions emerged in terms of cooking properties. The six AFLP Eco+3/Mse+3 different primer combinations applied in this research revealed different levels of polymorphism among the faba bean accessions and a total number of 346 amplicons were generated. Around 60% of amplicons displayed a polymorphic pattern among different accessions. Cluster analysis on morphological, technological, chemical and molecular data placed the all five ‘Larga di Leonforte’ accessions into a separated group, and the Sicilian material shares a fairly large amount of similarity with respect to the cultivar ‘Aguadulce Samba’ selected in Spain.     

Diversity characterisation of broccoli (Brassica oleracea L. var. italica Plenck) landraces for their on-farm (in situ) safeguard and use in breeding programs

Brassica oleracea L. is an important species that originated in the eastern Mediterranean area. From there, B. oleracea L. subsp. capitata (L.) DC. convar. botrytis (L.) Alef. var. italica Plenck (broccoli) was introduced into Italy where considerable diversification took place. Several landraces of broccoli, which are highly appreciated by local people for their organoleptic traits, are presently cultivated in both large fields and home gardens. In this study, 10 landraces, 4 synthetics derived from landraces and 4 of the most used F1 hybrids were characterised for morpho-physiological and molecular traits with the principal aim of feeding information into the landrace protection schemes being developed in Italy and into breeding programs. The landraces and derived synthetics are well differentiated from each other and from the F1 hybrids for important agronomic and genetic traits. The principal component analysis based on analysis of morpho-physiological traits showed that overall the 2 main components accounted for 67.74 % of the total variation. The neighbour joining tree, based on 23 microsatellite markers, grouped accessions into 2 main clusters. One includes one synthetic variety and 2 F1 hybrids, while the other cluster includes all the other accessions. There were also several sub clusters. Most of the materials examined showed within-accession diversity. The information gathered in this study can be used to protect landraces through the schemes presently being developed in Italy and to register them in the European common catalogue of “conservation varieties”. In situ (on-farm) conservation of landrace diversity is discussed with reference to landrace use.     

Genetic diversity of potato landraces from northwestern Argentina assessed with simple sequence repeats (SSRs)

Andean potato varieties are cultivated in the northwest of Argentina and constitute the most important staple food for the local farmers. The genetic diversity of 155 accessions conserved at the Genebank of Balcarce (INTA) was tested using four microsatellites. Three commercial potato varieties of Tuberosum group and one accession of Curtilobum group were used as outgroups. The presence of bands was scored for each microsatellite and for each accession and the data were analysed by principal coordinate analysis. The polymorphism information content was obtained for each molecular marker from banding patterns. Analysis of molecular variance was carried out with a variable number of accessions for each landrace, from different departments and sites within departments. More than one genotype was detected in the majority of the potato landraces. Some accessions within each landrace did not differentiate. AMOVA revealed that most of the genetic variation occurred among sites within departments and among local varieties. These findings are discussed considering the agricultural practices carried out in the Andean farming system.     

Geographical comparison of genetic diversity in Asian landrace wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) germplasm based on high-molecular-weight glutenin subunits

Glutenin largely determines wheat bread baking quality. As high-molecular-weight glutenin subunit (HMW-GS), related to Glu-1 loci, determines wheat flour elasticity, it correlates strongly with bread-making quality. This study was aimed at clarifying genetic variations in bread-making characteristics between East and West Asian wheat landrace germplasms, by investigating HMW-GS allelic composition of 1068 wheat accessions. Herein, the accession number having reported HMW-GS pattern in previous studies was 855. However, the accession number with newly detected HMW-GS patterns was 114. These new HMW-GS patterns were classified into 4 types based on similarity. Eight Korean accessions with these four types were identified. Concerning landrace germplasm nature, 99 accessions showed heterogeneous patterns caused by seed mixture. The Glu-1 loci allelic variation analysis, revealed that the percentages of Glu-A1c (73.6%), Glu-B1b (60.2%), and Glu-D1a (68.5%) were highest at Glu-A1, Glu-B1, and Glu-D1 loci, respectively. The incidence of preferable alleles for bread baking was high in Chinese accessions. In bread-making quality evaluation using Glu-1 score, 24 among 35 accessions with full score were from China. The polymorphic information content index of each origin based on HMW glutenin subunit combination showed that West Asian and neighboring-regional landraces, excluding Afghanistan ones, were more diverse than East Asian landraces excluding Chinese ones. Cluster analysis based on Glu-1 allelic combination showed that many Korean, Japanese, and Afghan accessions were in the same group. However, many Chinese and other West Asian accessions were in the other group despite geographical distance.     

Genetic diversity among Polish landraces of common oat (Avena sativa L.)

All over the world about 220,000 accessions of the genus Avena is preserved in gene banks. Polish oats collection, in the National Centre for Plant Genetic Resources, consists of about 2,500 accessions. More than 80 % of them belong to the species Avena sativa. Only 136 accessions have the landrace or traditional cultivar status and 91 of them were collected in Poland. The main objective of this study was to estimate genetic diversity of a set of 67 Polish landraces using inter simple sequence repeat (ISSR) markers. We also tried to determine whether genetic diversity depended on the region and altitude of collection site and if there was a relationship between DNA polymorphism and some morphological traits. The accessions were collected between years 1973 and 1999 from areas of Northern, Southern and Eastern Poland. The obtained results indicated a relatively low diversity (0.09–0.37) of the common oat landraces. Analysis of molecular variation (AMOVA) showed that there were no significant differences within designated geographic regions. It indicated that at the same time the altitude of collection site had significant effect on the genetic differentiation of the accessions. AMOVA also showed the presence of variance between groups formed on the basis of the colour of lemma. White lemma accessions were also characterized by highest genetic distinctiveness. Considering the evident impact of local climatic conditions on the genetic distinctiveness of Polish landraces utility of them in the Polish breeding programs should be considered.     

Patterns of SSR variation in bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) seeds under ex situ genebank storage and accelerated ageing

Maintaining seed viability and germplasm integrity is a challenging task in conservation of plant genetic resources, as seeds under storage will lose viability and genetic changes will occur. Attempt was made to analyze the patterns of genetic changes in wheat germplasm under ex situ genebank storage and accelerated ageing treatments. A set of 16 naturally aged wheat accessions under ex situ genebank storage since 1994 were sampled. Four recently regenerated wheat accessions were selected, four random seed samples were chosen from each accession, and three of them were exposed to three different accelerated ageing treatments. These 32 seed samples in two germplasm sets displayed a range of germination rates from 4 to 98 %. Thirty-seven microsatellite markers representing 21 wheat chromosomes were applied to screen 12 seeds of each sample and 449 SSR alleles were scored. Large SSR variation was found in each germplasm set. There was 73.1 % of the total SSR variation present among the naturally aged samples and 78.2 % present among the accelerated ageing samples. Several analyses for genetic association consistently revealed no clear genetic separations among samples of high or low germination rates in both germplasm sets. Samples under different accelerated ageing treatments did not show much genetic differentiations from the original sample of each accession. Mantel tests revealed non-significant associations between SSR variability and sample germination rates for both germplasm sets. These findings are useful for understanding seed deterioration under different ageing conditions and suggest that genome-wide SSR variability may not provide sensitive markers for the monitoring of wheat seed viability.     

Chromatographic fingerprinting of <Emphasis Type="Italic">Lupinus luteus</Emphasis> L. (Leguminosae) main secondary metabolites: a case of domestication affecting crop variability

Secondary metabolites (SMs), such as alkaloids and raffinose oligosaccharides (RFOs), play important roles in plant physiology. Although alkaloid and RFO phenotypic variation has been reported for yellow lupin (Lupinus luteus L.), most evaluations have used a reduced number of accessions; thus, limiting the understanding of accumulation patterns and variation ranges. The main goal of this research was to assess alkaloid and RFO content in a diverse L. luteus sample to understand possible SM accumulation patterns across this legume species. Eighteen yellow lupin accessions were analyzed using high performance thin layer chromatography to provide insights on seed and leaf RFO and alkaloid phenotypic variation. Co-dominant markers (170) were used to examine genetic relationships among L. luteus accessions and possible accumulation patterns across closely related genotypes. Significant differences were observed for seed and leaf RFOs. Total seed RFO accumulation ranged from 79.738 to 131.079 mg g^?1. Raffinose, stachyose, and verbascose were observed in all genotypes’ seeds, but at different RFO concentrations. Raffinose was the only RFO detected in leaves (2.793–0.4224 mg g^?1). Total alkaloid accumulation ranged from 0.22 to 15.12 and 0.00 to 8.007 mg g^?1 for seeds and leaves, respectively. Lupinine, sparteine, and gramine were observed in seeds and leaves, and showed a wide range of variation. Neighbor-Joining (NJ) analysis showed an apparent pattern of seed alkaloid accumulation, most likely due to domestication events. However, high RFO accumulating accessions were scattered across the NJ tree. Alkaloid and RFO significant phenotypic variation will not only help to understand the roles of these SMs in L. luteus, but also to uncover the genetic basis behind their accumulation.     

Genetic characterization of guava (Psidium guajava L.) germplasm in the United States using microsatellite markers

Genetic diversity of 35 Psidium guajava L. accessions and three related species (P. guineense Sw., P. sartorianum (O. Berg) Nied. and P. friedrichsthalianum (O. Berg) Nied.) maintained at the U.S. Department of Agriculture (USDA), National Plants Germplasm System, Hilo, HI, was characterized using 20 simple sequence repeat (SSR) markers. Diversity analysis detected a total of 178 alleles ranging from 4 to 16 alleles per locus. The observed mean heterozygosity (0.2) and inbreeding coefficient (0.8) indicated a high level of inbreeding among the accessions tested. Multi-locus DNA fingerprints based on the 20 SSR loci unambiguously differentiated all accessions and revealed the absence of duplicated samples. Ordination and cluster analyses suggested that the genetic relationships between majorities of the accessions could be explained by geographic origin, mainly including tropical America, Southeast Asia and Hawaii. A Bayesian cluster analysis partitioned the accessions into two groups and the partition was largely compatible with the result of ordination analyses. Distance-based cluster analyses further indicated that accessions from same geographical region or breeding programs grouped together in spite of the inter-regional exchange of germplasm. Accessions from Southeast Asia were dominantly white fleshed, whereas accessions from tropical America and Hawaii exhibited diverse flesh colors. The results also indicated that accessions from the same region were likely derived from a small number of common ancestors or progenitors. All 20 SSRs were transferable to P. guineense, P. sartorianum and P. friedrichsthalianum, indicating a close relationship between the cultigens and wild relatives. Application of SSR markers in the USDA/Agricultural Research Service germplasm collection provides new insight into the diversity of the guava germplasm, which will be valuable in future breeding endeavors and the conservation of guava genetic resources.     

Cytoplasmic diversity in Brassica rapa L. investigated by mitochondrial markers

Brassica rapa L. is an important vegetable and oilseed crop. Cytoplasmic diversity of 36 B. rapa accessions was analyzed using the mitochondria-specific markers. Twelve representative materials including five additional Brassica species and one Eruca sativa Mill. were used as references. A modified multiplex PCR amplification using four pairs of primers was performed to test the mitochondrial types (mitotypes) of the tested materials. Ten accessions were detected with Cam-I mitotype which could amplify 500 and 800 bp bands, twenty-two accessions with Cam-II mitotype which could amplify 500, 800 and 906 bp bands, one accession with Pol mitotype. Interestingly, three B. rapa accessions were revealed with nap mitotype, two of them were local landraces in northern Shaanxi, the third one was a variety from Gansu province which was developed using one local landrace from Northern Shaanxi as female parent. The considerable cytoplasmic diversity in B. rapa provides useful information on studying the possible origin and evolution of B. rapa accessions, and conservation of the germplasm.     

Genetic diversity in farmer grown spring barley material from Kyrgyzstan

Barley (Hordeum vulgare L.) is one of the economically important cereal crops grown in the highlands of the Kyrgyz Republic. In marginal agricultural areas of the country barley is considered as irreplaceable and very suitable fodder crop for livestock. In this study the genetic diversity of currently grown farmers’ spring barley material from Naryn and Issik-Kul provinces is described. In order to capture maximum diversity present in farmer field’s different morphological types were collected. The 22 spring barley accessions collected in 2008 were described morphologically and analysed using fourteen simple sequences repeat (SSR) markers. Most material was two-rowed barley, 78 %, whereas six-rowed barley was only 22 %. The spike colours present were light yellow, dark yellow, brown and violet. The material varied also for such characters as length of spike and number of kernels per spike. For the SSR loci 129 alleles were detected with an average of 9.3 alleles per locus and a genetic diversity with an average of h = 0.721 per locus. Based on analysis of molecular variance the 71.3 % of genetic diversity accounted for within accession variation, whereas 27.4 % for among accession variation. The variation between provinces was not significantly different. A group of two-rowed barley was identified that was more closely related to six-rowed barley than to the rest of the two-rowed barley material. Two-rowed material currently grown in Kyrgyzstan is more diverse than six-rowed barley (h = 0.65, A = 8.8 vs. h = 0.59, A = 3.1). From this study we could conclude that the materials nowadays grown by farmers are mixtures. Since there are no geographic differentiation between these mixtures suggests that materials is frequently inter-exchanged between farmers and are most probably not landrace material. The question concerning the origin of these mixtures remains unsolved and further investigation will be carried out to identify sources of variation in farmers’ field.