Effects of Neotyphodium fungi on Lolium multiflorum seed germination in relation to water availability.

BACKGROUND AND AIMS: Temperate endophyte-infected (Neotyphodium sp.) grasses have been shown to exhibit an ecological advantage over endophyte-uninfected grasses under abiotic stressful conditions. It is predicted that endophyte-infected plant populations will display higher rates of germination and proportion of germinated seeds under limiting water conditions. METHODS: The hydrotime regression model was used to describe the effect of Neotyphodium endophyte on seed germination of Lolium multiflorum at different water potentials. Additionally, seed mortality after water stress exposure was estimated in endophyte-infected and -uninfected seeds. KEY RESULTS: Endophyte infection inhibited seed germination at all water potentials. The hydrotime model described satisfactorily the germination responses, and revealed that endophyte-free seeds exhibited higher rates of and final percentage germination, probably due to a lower base water potential compared with endophyte-infected seeds. However, Neotyphodium endophyte conferred a higher rate of survival in those seeds that remained ungerminated when exposed to highly water stress conditions. CONCLUSIONS: Changes produced by Neotyphodium endophyte in L. multiflorum seeds might affect fitness in particular ecological scenarios. For example, the presence of the endophyte may curtail seed germination when water is limiting, reducing the risk of seedling death. Conversely, endophyte-free seeds would display an enhanced germination, ensuring a more rapid seedling establishment if later water conditions do not restrict plant growth.

Cloning and expression of a sorghum gene with homology to maize vp1. Its potential involvement in pre-harvest sprouting resistance.

Pre-harvest sprouting (PHS) in sorghum is related to the lack of a normal dormancy level during seed development and maturation. Based on previous evidence that seed dormancy in maize is controlled by the vp1 gene, we used a PCR-based approach to isolate two Sorghum bicolor genomic and cDNA clones from two genotypes exhibiting different PHS behaviour and sensitivity to abscisic acid (ABA). The two 699 amino acid predicted protein sequences differ in two residues at positions 341 (Gly or Cys within the repression domain) and 448 (Pro or Ser) and show over 80, 70 and 60% homology to maize, rice and oat VP1 proteins respectively. Expression analysis of the sorghum vp1 gene in the two lines shows a slightly higher level of vp1 mRNA in the embryos susceptible to PHS than in those resistant to PHS during embryogenesis. However, timing of expression was different between these genotypes during this developmental process. Whereas for the former the main peak of expression was observed at 20 days after pollination (DAP), the peak in the latter was found at later developmental stages when seed maturation was almost complete. Under favourable germination conditions and in the presence of fluridone (an inhibitor of ABA biosynthesis), sorghum vp1 mRNA showed to be consistently correlated with sensitivity to ABA but not with ABA content and dormancy.

The TATA-less promoter of VP1, a plant gene controlling seed germination.

Vp1 is a seed-specific gene involved in the control of dormancy and germination. We here present the complete sequence of the sorghum vp1 promoter/enhancer region highlighting its main features, especially the lack of canonical TATA and CAAT boxes and the presence of elements responsive to abscisic acid and light. The region closest to the start of transcription is highly homologous to the partial proximal sequence reported for the maize vp1 promoter. This region is interrupted by a 57-nt stretch containing 14 CT microsatellite repeats. We observed a poor overall homology to the promoter from abi3 gene, the Arabidopsis counterpart bearing a similar coding sequence. However, there exists a high degree of homology (89%) between a TATA-rich 103-bp stretch of the sorghum vp1 promoter located about 700 nt upstream of the startpoint and miniature inverted transposable elements (MITEs) interspersed within the sorghum seed-specific kafirin cluster. This sorghum MITE-like element displays considerable homology (68%) to the TATA-less promoter from the sorghum NADP-malate dehydrogenase gene and lesser similarity to the Tourist, Pilgrim and Batuta MITEs previously identified within the promoter from the maize Abp1 (auxin-binding protein) gene.

Hormonal Regulation of Dormancy in Developing Sorghum Seeds.

The role of abscisic acid (ABA) and gibberellic acid (GA) in determining the dormancy level of developing sorghum (Sorghum bicolor [L.] Moench.) seeds from varieties presenting contrasting preharvest sprouting behavior (Redland B2, susceptible; IS 9530, resistant) was investigated. Panicles from both varieties were sprayed soon after pollination with fluridone or paclobutrazol to inhibit ABA and GA synthesis, respectively. Fluridone application to the panicles increased germinability of Redland B2 immature caryopses, whereas early treatment with paclobutrazol completely inhibited germination of this variety during most of the developmental period. Incubating caryopses in the presence of 100 [mu]M GA4+7 overcame the inhibitory effect of paclobutrazol, but also stimulated germination of seeds from other treatments. IS 9530 caryopses presented germination indices close to zero until physiological maturity (44 d after pollination) in control and paclobutrazol-treated particles. However, fluridone-treated caryopses were released from dormancy earlier than control and paclobutrazol-treated caryopses. Incubation in the presence of GA4+7 stimulated germination of caryopses from all treatments. Our results support the proposition that a low dormancy level (which is related to a high preharvest sprouting susceptibility) is determined not only by a low embryonic sensitivity to ABA, but also by a high GA content or sensitivity.

Mineral allocation to reproduction in Sorhum bicolor and Sorghum halepense in relation to parental nutrient supply.

This experiment investigated the effect of parental nutrient shortage on the allocation of five nutrients to seeds and rhizomes in Sorghum halepense, a perennial, noxious weed, and to seeds in Sorghum bicolor, an annual, cultivated species. Plants from both species were grown from seeds and supplied with fertilizer at three concentrations. The allocation of biomass and nutrients (N, P, K, Ca and Mg) to reproductive and vegetative parts was determined. Relative biomass allocation to reproduction (either sexual or vegetative) remained constant in S. halepense in spite of large differences in total plant weight. In S. bicolor, however, biomass allocation to sexual reproductive structures decreased significantly with decreasing nutrient supply. Individual seed weight was not modified by parental nutrient supply in S. halepense, but it increased with decreasing nutrient availability in S. bicolor. Important differences in mineral allocation to seeds were found between the two species. While S. bicolor seeds were largely buffered from the differences in parental nutrient status, concentration of nutrients in S. halepense seeds decreased significantly with decreasing supply for all the nutrients analyzed except Ca. However, mineral nutrient concentration in S. halepense rhizomes remained remarkably constant despite differences in the external supply, evincing the priority given to vegetative reproduction at the expense of sexual reproduction. Overall, the pattern of nutrient allocation in S. bicolor seeds under different nutrient supply resembled the pattern observed in S. halepense rhizomes, but it had little resemblance to the pattern of nutrient allocation in S. halepense seeds. The results are discussed in terms of differences and similarities in the reproductive strategy of these two species.

Changes in germinability, ABA content and ABA embryonic sensitivity in developing seeds of Sorghum bicolor (L.) Moench. induced by water stress during grain filling.

The effect of intermittent water stress during grain filling on the germinability of developing seeds of S. bicolor was investigated. The drought treatment was imposed in cycles within the maturation period by withholding water for 5-6 days, rewatering at the end of each drought cycle and withholding water again. Changes in abscisic acid (ABA) content and embryonic sensitivity to ABA in the maturing seeds were also monitored in order to find out if there were any parallel changes with seed germinability resulting from drought conditions. Seeds developing in plants subjected to drought showed a high level of germinability earlier in the maturation period than did control seeds; consequently, they were less resistant to pre-harvest sprouting as shown when panicles were exposed to high humidity conditions. Very high levels of ABA accumulated in the early stages of development in seeds maturing on water-stressed mother plants; however. ABA content fell markedly when the seeds stopped growing, and remained significantly below those recorded in control seeds until the end of the maturation period. Development under drought conditions decreased the sensitivity of the isolated embryo to exogenous ABA by about 10-fold. The good agreement found between germinability, endogenous ABA concentrations and embryo sensitivity to ABA at different stages of development, suggests a key role for ABA as a major inhibitor of precocious germination and shows that changes in germinability caused by water stress during grain filling are likely to be related to changes in ABA pool size in the developing seed.