Seed viability of five wild Saudi Arabian species by germination and X-ray tests

Our objective was to evaluate the usefulness of the germination vs. the X-ray test in determining the initial viability of seeds of five wild species (Moringa peregrina, Abrus precatorius, Arthrocnemum macrostachyum, Acacia ehrenbergiana and Acacia tortilis) from Saudi Arabia. Usually several days were required to determine the viability of all five species via germination tests. However, X-ray test will give immediate results on filled/viable seeds. Seeds of all species, except Acacia ehrenbergiana and Acacia tortilis showed high viability in both germination (96–72% at 25/15 °C, 94–70% at 35/25 °C) and X-ray (100–80%) test. Furthermore, there was a general agreement between the germination (19%, 14% at 25/15 °C and 17% and 12% at 35/25 °C) and X-ray (8%, 4%) tests in which seed viability of Acacia ehrenbergiana and Acacia tortilis was very low due to insect damaged embryo as shown in X-ray analysis. Seeds of Abruspreca torius have physical dormancy, which was broken by scarification in concentrated sulfuric acid (10 min), and they exhibited high viability in both the germination (83% at 25/15 °C and 81% at 35/25 °C) and X-ray (96%) tests. Most of the nongerminated seeds of the five species except those of Acacia ehrenbergiana and Acacia tortilis, were alive as judged by the tetrazolium test (TZ). Thus, for the five species examined, the X-ray test was proved to be a good and rapid predictor of seed viability.     

Temporal,cell-specific,and tissue-preferential expression of myrosinase genes during embryo and seedling development in Sinapis alba

The expression of the genes for two types of myrosinase (EC 3.2.3.1), designated MA and MB, during embryo and seedling development was investigated in Sinapis alba L. by in-situ and RNA slot-blot analyses. The expression of MA and MB genes followed similar temporal profiles during embryogenesis, but MB mRNA was present in considerably higher amounts than MA mRNA. In the embryo, both MA and MB genes are activated in cotyledons and axis. The MB genes are preferentially expressed in the cotyledons whereas MA genes are preferentially expressed in the axis. In the developing seedling, MA mRNA was not present in the organs investigated. By contrast, MB mRNA was found in appreciable amounts in hypocotyls, cotyledons and developing leaves. The MB genes seem to be activated preferentially in tissues undergoing rapid cell division and — or cell expansion.Abbreviations DAP days after pollination - MA, MB A type, B type myrosinases in Sinapis alba Anna-Stina Höglund (Uppsala Genetic Center) is gratefully acknowledged for valuable discussion, Anders Gobl (Department of Immunology, Uppsala University) for kindly advice with the labeling of probes and Qingzhu Zhai (Department of Pharmaceutical Biosciences, Uppsala University) for help with seed harvest. This work was supported by grants from the Swedish Research Council for Forestry and Agriculture.     

Genetic variation of liverleaf (Hepatica nobilis Schreb.) in Bavaria against the background of seed transfer guidelines in forestry and restoration

The liver leaf, Hepatica nobilis (Ranunculaceae), is a perennial forest understory herb with specific habitat requirements, which occurs on calcareous soils in beech and oak forests. In Bavaria it can be found, therefore, only in four geographic regions, which are Franconia, the Franconian-Swabian Jura, the prealpine transitions region and the prealpine moraine belt with the Alps. In German forestry, provenance delineations are used since 1987. Similarly for herbs seed provenances and production areas are applied in commercial seed production and restoration. In this study we analyzed whether the genetic variation of H. nobilis reflects the geographic distribution of the species in Bavaria or the provenance delineations used for seed production.We applied AFLPs to study genetic variation within and between 24 populations of H. nobilis.Our analysis revealed high levels of genetic variation within and moderate variation between populations. Variation between seed production areas and seed provenances was low and comparable to the variation between geographic regions. Genetic variation within populations or rare fragments did not differ between production areas, provenances or geographic regions. A significant positive relation of genetic and geographic distances was present within 100 km.Pollination and seed dispersal seem mainly to happen within populations of the myrmecochorous H. nobilis, whereas long-distance dispersal is presumably occasional and random. Our study supports the relevance of dispersal traits for seed collections used in conservation. The protection of historically old beech and oak forests is exceedingly important to preserve the genetic variation of H. nobilis in Bavaria.     

Wetland drying indirectly influences plant community and seed bank diversity through soil pH

High altitude wetlands on the Tibetan Plateau have been shrinking due to anthropogenic disturbances and global climate change. However, the few studies that have been conducted on wetlands are inconclusive about the effect of soil moisture on seed banks and potential of seed banks in wetlands with different levels of soil moisture for regeneration of dried wetlands. We investigated seed banks and plant communities along a soil moisture gradient. A structural equation model was used to analyze the direct and indirect effects of soil moisture on seed banks, as well as the relationship between plant communities and seed banks. Although soil moisture had no direct effects on seed bank richness and density and indirect effects on seed banks through plant community, it had indirect effects on the seed bank through soil pH. Soil moisture also did not have direct effects on plant community richness, but it had indirect effects through soil pH. Plant community composition changed with soil moisture, but aboveground plant abundance and seed banks composition did not change. Low similarity exists between plant community and seed banks for all wetlands, and similarity decreased along the moisture gradient. The key factor determining plant community diversity was soil pH, while seed bank diversity was mainly affected by soil pH and plant community diversity with wetland drying. Although potential for regenerating the plant community from the seed bank decreased with an increase in soil moisture, drained wetlands still have enough residual seeds for successful restoration of species-rich alpine meadows.     

ABA对黑黄檀种子萌发的抑制作用以及其他植物激素对ABA的拮抗作用

以云南特有濒危树种黑黄檀(Dalbergia fusca)的种子为材料,研究了脱落酸(ABA)对种子萌发的抑制作用,以及种子萌发过程中吲哚乙酸(IAA)、赤霉酸(GA_3)、6-苄基腺嘌呤(6-BA)和乙烯利对ABA的拮抗作用.黑黄檀种子萌发的适宜温度为30℃.交替光照(14 h光照和10 h黑暗)以及黑暗对种子萌发没有明显的影响.0.001～0.1 mmol/L ABA不影响种子的萌发率,但降低种子的萌发进程;1 mmol/L和2.5mmol/L ABA显著地抑制种子的萌发率和萌发进程.种子的萌发率不被0.0001～1 mmol/L IAA和GA3、0.0001～0.1 mmol/L 6-BA、以及0.001～10 mmol/L 乙烯利(乙烯供体)的影响,但被1 mmol/L 6-BA抑制.1mmol/L ABA对种子萌发的抑制作用能被0.01～1 mmol/L IAA、0.01～1 mmol/L GA3、0.001～0.1 mmol/L 6-BA和0.1～10 mmol/L乙烯利所拮抗,而且这种拈扰作用与植物激素的类型和浓度有关.0.01 mmol/L 6-BA和0.1 mmol/L乙烯利对l mmol/L ABA抑制作用的拈抗不能被添加0.001 mmol/L IAA或者O.001 mmol/L GA3加成.但0.1 mmol/L 乙烯利对1 mmol/L ABA抑制作用的拮抗能够被添加O.01 mmol/L 6-BA或者0.1 mmol/L 6-BA加成,导致更高的萌发率和幼苗生长.     

Seed dispersal of Korean pine Pinus koraiensis labeled by two different tags in a northern temperate forest, northeast China

Two types of tagging methods, i.e., a 1 × 3-cm tin tag attached to seed with a 10- to 12-cm metal wire (total weight 0.32 g) and a 2 × 4-cm white plastic tag fastened to seed with an identical metal wire (total weight 0.57 g) were used to study their effects on seed dispersal of Korean pine by small rodents. A total of 600 seeds were released to assess four main points: (1) difference in seed survival rates, (2) difference in caching behaviors of small rodents, (3) difference in dispersal distances, and (4) proportion of seed missing. The results demonstrated that seed removal for wire-plastic-tagged seeds was faster than that for wire-tin-tagged seeds. There was no apparent difference in the proportion of seeds eaten in situ (42% and 52% for wire-plastic-tagged seeds and wire-tin-tagged seeds, respectively). We found 41% and 1% of seeds were moved and hoarded for wire-plastic-tagged seeds and wire-tin-tagged seeds, respectively. However, 2.33% and 14% of seeds were missing, and their ultimate fates were not known for wire-plastic-tagged seeds and wire-tin-tagged seeds, respectively. We found the wire-plastic-tagged seeds easier to track than the wire-tin-tagged seeds due to the fact that the white plastic tags were more salient than the tin tags in field environments. The average dispersal distances were 4.11 ± 2.40 m and 3.01 ± 2.06 m for wire-plastic-tagged seeds and wire-tin-tagged seeds, respectively, and showed great difference. Despite most being eaten in situ or after removal, 41% of seeds were cached for wire-plastic-tagged seeds, much more than for wire-tin-tagged seeds. A total of 71 primary caches (123 seeds) were found for wire-plastic-tagged seeds, with the average and maximum cache sizes being 1.73 and 6, respectively. However, only three caches were found, and cache size was equal to one for wire-tin-tagged seeds. The above data suggests there is some uncertainty in different tagging methods to used track seed fates. Despite their effectiveness in helping to trace seed dispersal or movement by seed-dispersing rodents, different tagging methods—including size, color, and mass—need to be fully understand in enclosure experiments .     

Development of a seed DNA-based genotyping system for marker-assisted selection in maize

Leaf collection from the field, labeling and tracking back to the source plants after genotyping are rate limiting steps in leaf DNA-based genotyping. In this study, an optimized genotyping method using endosperm DNA sampled from single maize seeds was developed, which can be used to replace leaf DNA-based genotyping for both genetic studies and breeding applications. A similar approach is likely to be suitable for all plants with relatively large seeds. Part of the endosperm was excised from imbibed maize seeds and DNA extracted in 96-tube plates using individuals from eight F₂ populations and seven inbreds. The quality of the resultant DNA was functionally comparable to DNA extracted from leaf tissue. Extraction from 30 mg of endosperm yields 3–10 μg DNA, which is sufficient for analysis of 200–400 agarose-gel PCR-based markers, with the potential for several million chip-based SNP marker analyses. By comparing endosperm DNA and leaf DNA for individuals from an F₂ population, genotyping errors caused by pericarp contamination and hetero-fertilization were found to average 3.8 and 0.6%, respectively. Endosperm sampling did not affect germination rates under controlled conditions, although under normal field conditions the germination rate, seedling establishment, and growth vigor were significantly lower than that of non-sampled controls for some genotypes. However, careful field management can compensate for these effects. Seed DNA-based genotyping lowered costs by 24.6% compared to leaf DNA-based genotyping due to reduced field plantings and labor costs. A substantial advantage of this approach is that it can be used to select desirable genotypes before planting. As such it provides an opportunity for dramatic improvements in the efficiency and selective gain of breeding systems based on optimum combinations of marker-assisted selection and phenotypic selection within and between generations.     

A new PCR-based marker on chromosome 4AL for resistance to pre-harvest sprouting in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Pre-harvest sprouting (PHS) is a complex trait controlled by multiple genes with strong interaction between environment and genotype that makes it difficult to select breeding materials by phenotypic assessment. One of the most important genes for pre-harvest sprouting resistance is consistently identified on the long arm of chromosome 4A. The 4AL PHS tolerance gene has therefore been targeted by Australian white-grained wheat breeders. A new robust PCR marker for the PHS QTL on wheat chromosome 4AL based on candidate genes search was developed in this study. The new marker was mapped on 4AL deletion bin 13-0.59-0.66 using 4AL deletion lines derived from Chinese Spring. This marker is located on 4AL between molecular markers Xbarc170 and Xwg622 in the doubled-haploid wheat population Cranbrook × Halberd. It was mapped between molecular markers Xbarc170 and Xgwm269 that have been previously shown to be closely linked to grain dormancy in the doubled haploid wheat population SW95-50213 × Cunningham and was co-located with Xgwm269 in population Janz × AUS1408. This marker offers an additional efficient tool for marker-assisted selection of dormancy for white-grained wheat breeding. Comparative analysis indicated that the wheat chromosome 4AL QTL for seed dormancy and PHS resistance is homologous with the barley QTL on chromosome 5HL controlling seed dormancy and PHS resistance. This marker will facilitate identification of the gene associated with the 4A QTL that controls a major component of grain dormancy and PHS resistance.     

Germination ecology of Central Asian Stipa spp: differences among species, seed provenances, and the importance of field studies

Stipa-species are wide-spread in Central Asia, but sexual reproduction in the dry steppes is rare. To facilitate conservation and restoration of these important rangelands, we studied germination characteristics of three common Mongolian Stipa-species under field- and lab conditions. Seeds of Stipa krylovii, Stipa gobica and Stipa glareosa were sown at the study site in Southern Mongolia over two consecutive years during which period tests were carried out to ascertain whether competition or herbivory are the main constraints of seedling establishment. In addition, we tested germination and seed viability in the laboratory under two different temperature regimes (20/10°C and 8/4°C), as well as the effect of cold-stratification. The lab experiments also included S. krylovii seeds originating from three climatically different provenances. None of the three Stipa-species seedlings emerged during the first 2 years of the field study. However, after an unusually intense rain event in the third year, 3% of S. krylovii, 0.6% of S. glareosa and 0.1% of S. gobica seeds germinated in the study plots. The factors ‘sowing-year’ and ‘vegetation’ significantly affected seedling emergence, whereas grazing had no effect at all. Under laboratory conditions a high percentage of viable seeds of S. gobica and S. glareosa germinated at both incubation temperatures, and cold-stratification had no effect on germination or viability. Germination of S. krylovii seeds required warmer temperatures and cold-stratification had a positive effect. Such evidence for dormancy was more pronounced in seeds from the moister, northern provenances. Germination of Stipa-species in the field is rare and only possible under exceptionally moist conditions. Conservation should thus concentrate on steppe conservation rather than on restoration. Where artificial reseeding is necessary, differences among species and also among different seed provenances should be taken into account.     

Seed bank spatial structure in semi-arid environments: beyond the patch-bare area dichotomy

The prevalence of patchy structures in vegetation is a common feature in semi-arid ecosystems. Although the effect of patches on seed density is widely known, we still lack information on how patch features affect seed bank density and composition. Our aim was to answer two basic questions: (1) How do seed bank density and composition vary within and outside patch aboveground physical limits? and (2) Do patch characteristics affect soil seed bank density and composition? We sampled 50 shrub patches in a semi-arid gypsum system in Central Spain, measuring patch size, composition and structure, and seed bank at three locations per shrub (centre, edge and outside). We calculated the effect of interior patch location, patch composition and structure on seed density and composition. Patches acted both as seed sources, increasing seed density in neighbouring areas and as seed sinks by trapping seeds from bare areas. Patch structure (erect perennial cover) had the greatest effect on seed bank density, whereas patch size and microslope had the greatest influence on bare area density. Patch structure, composition and interior location explained the variation in seed bank composition. Patch effect extends to the surrounding bare matrix creating a seed bank gradient in density and composition. This effect is modulated by patch structure and composition and affects seed bank composition. Our results suggest that the spatial structure of gypsum community seed banks may act as a mechanism for a spatial storage effect contributing to the maintenance of high levels of diversity in semi-arid environments