Effects of pollination distance on reproduction and offspring performance in Hypochoeris radicata: Experiments with plants from three European regions

Fragmentation of habitats has resulted in increased inbreeding for many plant species, while the introduction of foreign seed material for ecological restoration has resulted in crosses between plants from distant populations. Both processes may reduce plant fitness and increase the risk of extinction. Variation in the expression of inbreeding and outbreeding depression has been found among different genotypes and among populations, but little is known about large scale geographical patterns within species. We studied the effects of cross-proximity on seed production and offspring performance in the perennial meadow plant Hypochoeris radicata (Asteraceae) from five populations in each of three European regions (Bohemia in northwest Czechia, Northern Hesse in central Germany, and Salland in the central Netherlands). Five artificial cross types were conducted with varying proximity of mates: selfing (self), within family crosses (WF), within population crosses (WP), between population crosses (BP), and between region crosses (BR), and the offspring were grown in a common garden. Independent of the region of origin of the maternal plant, selfing, WF and BP crosses resulted in lower seed set and germination than WP crosses, indicating partial self-incompatibility, inbreeding depression and reduced performance in the F1 progeny resulting from outbreeding. However, crosses between regions resulted in similar seed set and germination as within population crosses. For late traits, the effects of inbreeding and interpopulation crosses differed among regions. WP crosses exhibited the highest survival, flowering and multiplicative fitness only in progeny from Czech maternal plants. Our results suggest that the sensitivity of populations to introgression may vary among regions and that outbreeding depression does not necessarily increase with interpopulation distance. However, the current study investigated only effects in the F1 in a common garden and outbreeding depression may be stronger in the F2 and in field populations.     

Morphological patterns in a world collection of <Emphasis Type="Italic">Cleome gynandra</Emphasis>

Cleome gynandra L. is a leafy vegetable native to sub-Saharan Africa and Asia and cultivated on dry areas. The plant plays an important role in the food and nutrition security of local communities. The objective of this study was to illustrate variation and diversity in the Cleome collection held at the World Vegetable Center. In total, 242 accessions were cultivated and morphologically characterized and analyzed. The gynophore and filament, both important organs of the flower, played a key role in taxonomical identification. High morphological variation was detected in traits including plant height, pod length, leaf size, flower color, and earliness. Distinct morphological differentiation was detected between Asian and African accessions. On average, the African accessions were larger, less uniform and later in flowering and seed maturation than the Asian accessions. The results were used to establish a core collection of 49 Cleome accessions. Gaps in the current collection were detected. High within-accessions diversity challenges the existing ex situ conservation system; to maintain diversity, seed should be collected from a large number of plants, both during the collection mission and in ex situ regeneration. The results are relevant for germplasm collection strategies and regeneration protocols for good genebank practices.     

Detection of genetic integrity of conserved maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) germplasm in genebanks using SNP markers

Twenty maize landrace accessions regenerated and conserved in five maize genebanks were investigated for genetic integrity using 1,150 Single Nucleotide Polymorphisms (SNPs) and 235 SNP haplotypes. The genetic diversity of three accessions changed significantly in terms of the average number of alleles per locus. Ten out of twenty accessions had significantly different SNP allelic frequencies, either after regeneration or in the same accession held in different genebanks. The proportion of loci with significant changes in SNP allelic frequency was very low (37/1,150). Changes in the major allelic frequency (MAF) for the majority of SNP loci (60.2–75.2%) were less than 0.05. For SNP haplotypes, the genetic diversity of four accessions changed significantly in terms of average number of haplotype alleles and polymorphic information content (PIC) per locus. The proportion of SNP haplotype alleles lost in the later generations ranged between 0 and 22.6%, and at the same time 0–19.9% of the SNP haplotype alleles appeared in later generations, however, these were absent in the earlier generations. Dynamic changes in genetic integrity, in terms of presence and absence of genes (alleles), by both SNP and SNP haplotype analysis were detected during regeneration. A suboptimum number of ears harvested in one generation can be combined with those from another, repeated regeneration to capture the diversity of the previous generation. Use of molecular markers during regeneration of accessions can help in understanding the extent of genetic integrity of the maize accessions in ex situ genebanks and in recommending the best practice for maintaining the original genetic diversity of the genebank accessions.     

Evaluation of yield and agronomic traits and their genetic variation in 488 global collections of Brassica napus L.

It is important to evaluate yield and agronomic traits when selecting for elite lines in Brassica napus L. In this study, 488 global collections of B. napus L. were evaluated for yield and agronomic traits during two consecutive years under growing conditions in central China. A series of phenotypic data for yield and agronomic traits were obtained. Large variations in yield and agronomic traits were found among these accessions, making selection possible for these characters. Chinese accessions, in general, tended to have higher plot yield (POY), higher seed yield per plant (PY), higher thousand-seed weight (TSW), more seeds per silique (SS), and fewer siliques per plant (PS) than foreign accessions. Comparing winter, semi-winter, and spring accessions, semi-winter accessions tended to have the highest POY, highest PY, highest TSW, most SS and fewest PS. Genotypic variation for TSW accounted for 71.23 % of the total variation, and it was the highest for all 12 of the phenotypic traits. Significant correlations were observed between different traits. Principal component analysis (PCA) revealed that significant variation among the traits with the first four principal components could explain 49.8 % of the total variation. Three major groups (winter, spring and semi-winter oilseed rape accessions) could be distinguished when projecting the accessions onto the first two PCAs. The information on variations in yield and agronomic traits detected in this study provided useful parents for rapeseed breeding. Moreover, the phenotypic data on yield and agronomic traits obtained may be used in our subsequent genome-wide association studies for B. napus.     

Morphological and reproductive characterization of guar (<Emphasis Type="Italic">Cyamopsis tetragonoloba</Emphasis>) genetic resources regenerated in Georgia,USA

Guar, Cyamopsis tetragonoloba is used worldwide for food stabilization, fiber source, food, and industry. The United States Department of Agriculture, Agricultural Research Service, Plant Genetic Resources Conservation Unit (USDA, ARS, PGRCU) conserves 1,298 accessions originating from India, Pakistan, and breeding lines from the USA. My objectives were to determine: (1) if these guar accessions can successfully be regenerated in Georgia, USA, (2) which traits contribute the most variation, and (3) to provide estimates for genetic relatedness among these accessions. Guar accessions were directly seeded to the field in Griffin, GA between early May and mid June 2006–2008. At 50% maturity, 73 accessions were characterized for morphological, phenological, and reproductive traits during the regeneration cycles. High quality plants regenerated from most of the accessions and produced 80 to more than 9,300 seeds per accession. Guar can be successfully grown and regenerated in Griffin, GA. Coefficients of variation and principal component analysis revealed variability among accessions for these traits evaluated. Cluster analysis separated guar accessions into three groups (clusters) based on low, intermediate or high seed numbers. Guar accessions clustered in groups 2 and 3 appear to be more genetically related than those in group 1.     

Characterization of sesame (<Emphasis Type="Italic">Sesamum indicum</Emphasis> L.) germplasm regenerated in Georgia,USA

Sesame, Sesamum indicum L. is used worldwide for cooking oil and food. The United States Department of Agriculture, Agricultural Research Service, Plant Genetic Resources Conservation Unit (USDA, ARS, PGRCU) conserves 1,226 accessions originating from Europe, Africa, Asia, North America, and South America. Sesame accessions were directly seeded to the field in Griffin, GA between 15 May and 01 June 2003–2007. At 50% maturity, 192 accessions were characterized for morphological, phenological, and reproductive traits during the regeneration cycles. High quality plants regenerated from all accessions produced 131 to more than 80,000 seeds per accession. Sesame can be successfully grown and regenerated in Griffin, GA. Coefficients of variation and principal component analysis revealed considerable variability among accessions for phenological, morphological and reproductive traits. Assuming appropriate levels of heritability, sufficient variation among sesame accessions exists for days to maturity, plant height, and reproductive traits among these sesame accessions to warrant breeding programs for new sesame cultivar development.     

Preservation of seed viability during 25 years of storage under standard genebank conditions

Maintaining sufficient viability is critical to the sustainability of ex situ conserved seed collections. For this reason, accessions are regenerated when viability falls below a predefined threshold. Viability is monitored by determining the germination ability of accessions at predefined time intervals. Optimizing the frequency of these germination tests, in order to avoid waste of resources, is hampered by the scarce availability of data about seed longevity, particularly for material maintained under genebank conditions. Here we report on the analysis of nearly 40,000 germination test results collected for a wide range of crop species over a 25-years period by the centre for genetic resources, the Netherlands (CGN), where seeds of genebank accessions are dried to 3–7 % moisture content and stored for the long term under near-vacuum in aluminium foil bags at ?20 °C. The results indicate that seed viability is well maintained for the large majority of seed lots during the first 25 years after regeneration as only 3.3 % of the monitoring tests revealed below-threshold germination values. It is argued that the majority of these sub-standard seed lots are due to other causes than seed ageing, including dormancy problems and estimation error in germination testing. For material, maintained under the seed management procedures and storage conditions practiced by CGN, it is therefore recommended to delay the first germination monitoring tests to 25 years after regeneration.     

Relationships in the <Emphasis Type="Italic">Diplotaxis–Erucastrum–Brassica</Emphasis> complex (Brassicaceae) evaluated from isoenzymatic profiles of the accessions as a whole. Applications for characterisation of phytogenetic resources preserved <Emphasis Type="Italic">ex situ</Emphasis>

Electrophoresis has become one of the most usual tools for plant studies. However, most analyses, using isoenzymes, have been carried out from a population geneticist’s point of view for the study of allelic frequencies. In present work, isoenzyme electrophoresis of bulked seeds was used as a rapid strategy for obtaining the electrophoretic profiles of accessions of a seed bank. The relationships amongst 30 taxa of the complex Diplotaxis-Erucastrum-Brassica (Brassicaceae family, tribe Brassicinae) were analysed, with the electrophoretic profiles of six isoenzyme systems. Each sample presented a stable and characteristic isoenzymatic pattern representative of the whole population. The enzyme markers provided 147 reliable bands which were scored as multistate traits (not with allelic value). UPGMA clustering method showed a dendrogram with three main groups and subgroups of samples that are in agreement with the recognised taxonomy of the taxa. This approach can be a rapid and efficient method to establish the specific enzyme patterns of conserved species populations of a germplasm bank, and the information could be included in the a database as part of documentation system.     

冷诱导基因CBF-4转化辣椒与转基因植株抗寒性分析

以4个辣椒（Capsicum annuum L. ）品种为试材,利用根癌农杆菌介导的带柄子叶转化法将拟南芥CBF-4基因转入辣椒,通过调整预培养、共培养以及延迟培养时间等因素,优化并完善了甜椒品种B12的遗传转化体系,并对获得的60株卡那霉素抗性再生植株进行PCR和RT-PCR检测表明,46株为阳性转基因植株,转化率高达76.67%。经过低温处理后,对阳性植株的抗寒性生理、形态指标分析发现,转基因植株的SOD、POD和MDA活性都明显高于未转基因植株,转基因植株比未转基因植株的株型紧凑,叶片肥厚、平直,叶色浓绿,叶面积大。     

Reduced seed production, inbreeding, and pollen shortage in a small population of a threatened tree, Magnolia stellata

We investigated how genetic factors and pollen shortage affect seed production in a small population of Magnolia stellata, a threatened insect-pollinated tree. We used microsatellite and manual pollination techniques, and compared a small population to a large one. Compared to the large population, the small population showed low allelic variation and had an F_IS significantly greater than 0 in adults, indicating that genetic deterioration, including genetic drift and inbreeding, may have occurred in adults. Manual self-pollination lowered seed production relative to manual cross-pollination to a different extent between populations: δ (the magnitude of inbreeding depression due to self-fertilization) was lower in the small population than in the large population. However, under natural pollination, the estimated embryo mortality rates after ovules self-fertilized were similar between the two populations because the primary selfing rate was higher in the small population. The ovule mortality rate due to pollen shortage and that due to factors independent of pollen shortage and selfing were both approximately 10% higher in the small population, suggesting that pollen transfer may have decreased and genetic deterioration effects may have increased in that population. These factors reduced seed production in the small population (female reproductive success = 0.3%) compared to the large population (2.6%). Our results suggest that seed production in the small population of M. stellata is strictly limited by elevated pollen shortage, selfing, and genetic deterioration in adults, which accelerate the risk of extinction.     

Breeding system and inbreeding depression in the clonal plant species Knautia arvensis (Dipsacaceae): implications for survival in abandoned grassland

Larger clonal sizes observed in abandoned than in managed grassland could result in increased geitonogamy in Knautia arvensis. The species was investigated for self-compatibility, seed set without pollinators, and inbreeding depression in a greenhouse experiment. Seed set was equivalent in hand-selfed and hand-outcrossed plants (c. 53%), indicating full self-compatibility. Seed set was low (2.8%) in caged, unmanipulated flower heads. There was no significant difference in seed weight, germination, and survival between selfed and outcrossed progeny. Juvenile biomass was significantly lower in selfed than in outcrossed progeny (on average 45% lower). Cumulative fitness calculations for germination, survival, and juvenile biomass showed that the mean inbreeding depression was 58% in offspring produced by selfing. The results suggest that in K. arvensis, which is protected against self-pollination within flower heads through protandry, the likely mechanism for selfing is via geitonogamy among flower heads. Geitonogamy among ramets within the larger clones observed in abandoned grassland could reduce offspring viability in K. arvensis since the fitness of selfed offspring was low. Inbreeding depression could therefore add to the recruitment problems experienced by plants in abandoned grassland, both in K. arvensis and probably in other clonal grassland species, depending on their growth structure and breeding system.     

Characterization of butterfly pea (<Emphasis Type="Italic">Clitoria ternatea</Emphasis> L.) accessions for morphology,phenology, reproduction and potential nutraceutical,pharmaceutical trait utilization

Butterfly pea, Clitoria ternatea is used in Africa as a companion crop and in the United States as an ornamental. The USDA, ARS, PGRCU curates 28 butterfly pea accessions. Butterfly pea accessions were transplanted from about 30-day-old seedlings to the field in Griffin, GA around 01 June 1999, 2003, 2006–2007 or directly sown in 2001. At 50% maturity, 19 accessions were characterized for morphology, phenology, and evaluated for regeneration. High quality plants regenerated from all accessions produced 6 to more than 3,400 total seeds. Butterfly pea can be successfully grown and regenerated in Griffin, GA. Coefficients of variation and principal component analysis revealed considerable variability among accessions for morphological and reproductive traits. Butterfly pea has potential to be used as a nutraceutical, pharmaceutical, or food. The flavonoid quercetin has been shown to reduce upper respiratory infections in humans while delphinidin and malvidin identified in butterfly pea flowers may inhibit various forms of cancer.     

Hypericum perforatum L. Seed Germination: Interpopulation Variationand Effect of Light, Temperature, Presowing Treatments and Seed Desiccation

Germplasm conservation of medicinal plants is of increasing interest and, when possible, seed banking is the most efficient system for ex situ conservation of these plant genetic resources. Hypericum perforatum L. (St. John's wort, Guttiferae) is a medicinal plant with evidence of efficacy as an anti-depressant. The aim of this work was to increase knowledge of its seed germination behaviour by studying 68 wild populations. Seed germination tests were carried out at 25/15 °C under a photoperiod of 16-h light/8-h darkness. Final germination percentages were highly variable depending on the accession, ranging from 6 to 98%. Similarly, germination rate (as expressed by T ₅₀ values) varied significantly from 6.1 to 23.0 days. The effect of seed desiccation with silica gel on subsequent germination was also studied. The effect of two other incubation temperatures (15 and 25 °C) and light (photoperiod or darkness) on seed germination was studied in several accessions. Temperature had no significant effect on final germination percentages. However, light significantly increased the germination of most but not all accessions assayed. Seeds from four accessions with low germination percentages were subjected to different presowing treatments that could increase germination: dry heat, hot water and gibberellic acid. Germination was promoted significantly by gibberellic acid in two of the four accessions assayed, but the thermal treatments did not enhance significantly the germination percentages. This study reveals that conclusions based on one population of Hypericum perforatum cannot characterize the germination behaviour for the entire species.     

Autofertility in a pure line faba bean (Vicia faba L.) germplasm collection

Summary A pure line faba bean germplasm collection (BPL) was derived by a process of cyclic single plant progenies starting in 1979 through 1985. In 1985/86, 840 of these pure line accessions from 677 original populations from 41 countries were evaluated for autofertility by comparing the mean of five plants where all flowers were tripped vs. five plants with no tripped flowers for number of seeds produced (seed index). In a replicated trial grown at the same time, seed yield, 100 seed weight, number of pods per plant, seeds per pod, and number of flowers per inflorescence were also evaluated. A high number of BPL accessions failed to set seeds in the screen houses without tripping, the mean seed index measure of autofertility was 0.51 with a C.V. of 74.2%. There were marked differences among countries of origin for autofertility, with Egypt having the highest (SI = 0.94) and the USSR the lowest (SI = 0.26) autofertility, however, most countries expressed a large range, suggesting that breeders should first look in their own material for this trait rather than searching in exotic germplasm. In general, there was a higher level of autofertility in Middle Eastern countries that may be due to natural selection for non-dependency on pollinators for high yield. Contrary to the findings of others, large seeded accessions had a high autofertility (SI = 0.704) compared to small seeded accessions (SI = 0.384), however, both had similar ranges. In general, there was no significant correlations among autofertility and yield and its components. The variability found in this large pure line collection for autofertility is of potential use for developing cultivars not dependent on insect populations for high yield and for developing self-pollinated faba bean.     

Swordbean (Canavalia ensiformis (L.) DC.) Genetic Resources Regenerated for Potential Medical, Nutraceutical and Agricultural Traits

Swordbean, Canavalia ensiformis has been used in Asia and Japan as a vegetable while in the United States sword bean is used as a source of the lectin concanavalin A. The USDA, ARS, PGRCU maintains 27 swordbean accessions. Swordbean accessions were transplanted from approximately 45-day-old seedlings to the field in Griffin, GA around 01 June from 1998 to 2003 (except 2001). After 3–4 months, 21 accessions were characterized for morphological traits and evaluated for regeneration. High quality plants regenerated from all accessions produced 19 to more than 1100 total seeds. Swordbean can be successfully grown and regenerated in Griffin, GA. Swordbean has potential to be used in numerous other ways including nutraceuticals, phytopharmaceuticals, and other agricultural products. Flavonoids such as rutin identified in swordbean seeds may decrease precancerous lesions in the large intestine while the phytochemical, beta-aminopropionitrile inhibits breast adenocarcinoma growth. Canatoxin, the potential pesticide from swordbean seeds showed toxicity to the cotton pest, Dysdercus peruvianus.     

Diversity in Indian sesame collection and stratification of germplasm accessions in different diversity groups

Sesame (Sesamum indicum L.) is one of the ancient oil crops, grown in India since over 5,000 years ago. Diversity in the Indian sesame collection (3,129 accessions), representing all eco-geographical regions, for a range of morphological and agronomic characters was studied. Wide variation in plant habit (plant height and branching pattern), pubescence of various plant parts (stem, leaf, corolla and capsule), flower colour and number of flowers per leaf axil, capsule characteristics (shape, size, number in the axil of a leaf and number of locules in a capsule), number of capsules per plant, number of seeds per capsule, mean seed weight, and yield per plant was recorded. A detailed multivariate analysis was performed on a set of 100 selected accessions representing different agro-ecological zones. The accessions were classified into 7 discrete clusters. The principal components analysis described the spatial relationship among the entities and confirmed groupings obtained through clustering. Based on the clustering pattern of 100 accessions, the entire collection was allocated to different clusters. Representation of various zones in 7 clusters gives us the opportunity to form distinct diversity groups making combined use of passport and characterisation data. These diversity groups would subsequently be used for sampling the accessions for building up a core collection of sesame, a project already operative at the NBPGR.     

小孢子培养获得松花型花椰菜DH再生植株

对5个松花型花椰菜(Brassica oleraceavar.botrytisL.)杂种一代的小孢子培养研究表明,小孢子胎胚发生主要依赖于基因型,庆农65天的每花蕾胚状体产量最高,平均达15.5个。松花菜的胚状体萌发率一般在30%左右,并有效地获得了大量的DH再生植株。冷激预处理能显著地影响花椰菜小孢子的胚胎发生,但供体材料间存在不同的结果。结球期和开花结角期再生植株的生育期与育性出现较大分离,可育且能正常结角的比例约占全部小孢子再生植株的50%以上,因而不再需要加倍处理。     

Seed banks in plant conservation: Case study of Santa Cruz tarplant restoration

Although conservation and restoration practitioners have focused on maximizing aboveground population size and seed set of rare plants, a clear understanding of seed bank dynamics is crucial to managing these species. Santa Cruz tarplant (Holocarpha macradenia) is a threatened annual forb restricted to coastal prairie habitats in central California. Holocarpha produces disk achenes germinating within a year of production and ray achenes forming a persistent seed bank. We constructed both deterministic and stochastic demographic models for a restored Holocarpha population, using demographic rates measured separately for unmanipulated plants and plants growing in plots where vegetation was clipped. The deterministic models indicated that regardless of germination from the seed bank, the population would decline without clipping or similar treatments that enhance survival and reproductive output. Deterministic models showed only a slight positive effect of increased ray seed germination rates on population growth, which would need to be balanced against a potential loss of buffering against environmental variation as the dormant seed bank was reduced. Our stochastic simulations suggested that extinction risk for Holocarpha populations would be minimized by intermediate levels of ray seed germination. Thus, managers should focus on improving the performance of aboveground plants before considering actions to stimulate germination, since the former will yield a greater increase in deterministic population growth and not sacrifice any buffering effect of the seed bank. This case study emphasizes the importance of considering dormant seeds and seed banks in designing successful restoration and management strategies for plant species at risk of extinction.     

小孢子培养有效获得松花型花椰菜DH再生植株

对5个松花型花椰菜杂种一代的小孢子培养研究表明，小孢子胎胚发生主要依赖于基因型，庆农65天的每花蕾胚状体产量最高，平均达15.5个。松花菜的胚状体萌发率一般在30%左右，并有效获得了大量的DH再生植株。冷击预处理能显著影响花椰菜小孢子的胚胎发生，但供体材料间存在不同的结果。结球期和开花结角期再生植株的生育期与育性出现较大分离，可育且能正常结角的比例约占全部小孢子再生植株的50%以上，因而不再需要加倍处理。     

基于适合度的普通群体基因型熵的逐代演变规律

本文在引入＂熵＂来定量普通群体的基因型的＂杂＂中,考虑到适合度的差异。把适合度量化为一个具有不同基因型群体产生的下一代达到性成熟的相对比例。推算出兼有自花与异花传粉群体的某世代中活到性成熟的3种基因型所占的比例。通过对多种不同的适合度及自交率的群体的不同世代基因型熵变曲线及平衡趋势分析,发现自交率越高,平衡状态的熵越小,达到平衡状态的速度越快。在自交率很高的情况下,变化过程的熵与平衡状态的熵很接近。