Seed Science Research

Induction of nine-cis-epoxycarotenoid dioxygenase 6 (NCED6), an abscisic acid (ABA) biosynthesis gene, alone is sufficient to suspend germination in testa-ruptured seeds, which are at the final step of germination. Molecular consequences of NCED6 induction in imbibed seeds were investigated by RNA sequencing. The analysis identified many unknown and uncharacterized genes that were up-regulated by NCED6 induction, in addition to the major regulators of ABA signalling. Interestingly, other NCEDs were up-regulated by NCED6 induction, suggesting that the major rate-limiting enzymes in the ABA biosynthesis pathway are subject to positive-feedback regulation. ZEAXANTHIN EPOXIDASE and ABSCISIC ALDEHYDE OXIDASE3, which function upstream and downstream of NCED, were also up-regulated in seeds by NCED6 induction, which suggests that the distinct layers of positive feedback loops are coordinately operating in the NCED6-induced seeds. SOMNUS (SOM), which was also up-regulated by NCED6 induction, was the major mediator of enhanced ABA signalling in NCED6-induced seeds. SOM exerted negative effects on GA biosynthesis, which also contributes to a high ABA–GA ratio and reinforces the suppressive state of germination. Besides these coding genes, long intergenic non-coding RNAs (lincRNAs) were also up-regulated upon NCED6 induction (termed N6LINCRs). Conditional expression of N6LINCR1 altered gene expression profiles in seeds. Twenty-six genes were up-regulated and 66 genes were down-regulated by the induction of N6LINCR1. These results suggest that some of N6LINCRs have a regulatory role in gene expression in seeds, which potentially contributes to the regulation of germination by ABA.

In the present study a combination of BLAST mediated homology search and 3′ RACE was utilized to isolate the full-length gene of Eleusine coracana alpha prolamin (Ec-α-prolamin) from finger millet. Phylogenetic analysis of Ec-α-prolamin along with related prolamin genes of different cereals and millets shows the clustering of Ec-α-prolamin in a separate group. Secondary structure prediction reveals 59.4% alpha helix structure, a structural hallmark of Ec-α-prolamin. Besides this, the protein also possesses a balanced proportion of all essential amino acids. Expression analysis based on qPCR shows increased accumulation of α-prolamin transcripts in developing grains of finger millet until attainment of seed maturity. Western blotting, using a monospecific anti-α-prolamin antibody, further confirmed the expression of a 22 kDa band in the S3 and S4 stages of developing spikes. The heterologous expression of isolated full-length Ec-α-prolamin could be potentially harnessed for making nutritionally enhanced functional food products and value-added industrial products.

Vitamin E (tocols) is a key metabolite for efficient scavenging of lipid peroxy radicals that cause membrane breakdown during seed ageing. However, in rice, this hypothesis has been tested for very few lines only and without considering intraspecific variation in genomic structure. Here, we present a correlation study between tocols and seed longevity using a diverse rice panel. Seeds of 20 rice accessions held in the International Rice Genebank at the International Rice Research Institute, representing Aus, Basmati/Sadri, Indica, temperate Japonica and tropical Japonica variety groups, were used for tocols analysis (quantification of α-, β-, γ- and δ-tocopherol/tocotrienol by ultra-performance liquid chromatography) and storage experiments at 45°C and 10.9% seed moisture content. To examine the effects of DNA-haplotype on the phenotype, the 700 K high-density single-nucleotide polymorphism marker dataset was utilized. Both seed longevity (time for viability to fall to 50%; p 50) and tocols content varied across variety groups related to the heterogeneity in the genetic architecture. Among eight types of tocol homologues, α-tocopherol and γ-tocotrienol were significantly correlated with p 50 (negatively and positively, respectively). While temperate Japonica varieties were most abundant in α-tocopherol, Indica varieties recorded 1.3- to 1.7-fold higher γ-tocotrienol than those of other groups. We conclude that the specific ratio of tocol homologues rather than total tocols content plays an important role in the seed longevity mechanism.

Post-harvest drying prolongs seed survival in air-dry storage; previous research has shown a benefit of drying moist rice seeds at temperatures greater than recommended for genebanks (5–20°C). The aim of this study was to determine whether there is a temperature limit for safely drying rice seeds, and to explore whether the benefit to longevity is caused by high-temperature stress or continued seed development. Seeds of two rice varieties were harvested at different stages of development and dried initially either over silica gel, or intermittently (8 h day–1) or continuously (24 h day–1) over MgCl2 at temperatures between 15 and 60°C for up to 3 days. Seeds dried more rapidly the warmer the temperature. Subsequent seed longevity in hermetic storage (45°C and 10.9% moisture content) was substantially improved by increase in drying temperature up to 45°C in both cultivars, and also with further increase from 45 to 60°C in cv. ‘Macassane’. The benefit of high-temperature drying to subsequent longevity tended to diminish the later the stage of development at seed harvest. Intermittent or continuous drying at high temperatures provided broadly similar improvements to longevity, but with the greatest improvements detected in a few treatment combinations with continuous drying. Heated-air drying of rice seeds harvested before maturity improved their subsequent storage longevity by more than that which occurred during subsequent development in planta, which may have resulted from the triggering of protection mechanisms in response to high-temperature stress.

The water uptake by the seed in arid and semi-arid ecosystems may not be continuous, but may occur in hydration and dehydration cycles (HD). In order to investigate the benefits of discontinuous hydration on seed germination of Pilosocereus catingicola subsp. salvadorensis subjected to environmental stresses, an imbibition curve was determined and times X, Y and Z selected, corresponding to 25, 50 and 75% of the first phase of imbibition, respectively. Seeds of two populations (Caatinga and Restinga) were subjected to 0, 1, 2 and 3 HD cycles, with 48 h of dehydration. Water and saline stress responses were tested at five osmotic potentials (0.0, –0.1, –0.3, –0.6 and –0.9 MPa). Discontinuous hydration provided greater tolerance to water and saline stress in the seeds from the Caatinga region. When the seeds were subjected to water stress, the reduction of the mean germination time (MGT) provided by HD cycles decreased with increasing concentration of the solution. In saline stress, the reduction of MGT was greater in the solutions with the highest concentrations. We conclude that the seeds of the same species collected from populations located in different ecosystems have different germination responses after passage through discontinuous hydration and HD cycles. This provides greater tolerance to environmental stresses but with different responses among populations.

The pattern of seed dispersal in time and space can affect plant fitness and the soil seed bank, and thus information is needed on this aspect of the seed biology of a species before it is selected for use in habitat restoration projects. Zygophyllum xanthoxylon is a super-xerophilous shrub that is a potential pioneer species for use in revegetating highly disturbed areas of the cold deserts of northwest China. We studied fruit release and soil seed banks of Z. xanthoxylon for 3 years in two cold desert habitats characterized by different degrees of drought and wind velocity. In our study, fruit (a three-winged capsule) release began in summer (June 2010, August 2011, July 2012) and extended for 9–10 months, but plants can be found in the population with previous- and current-year fruits attached to them. More than 50% of the fruits were released in the first 3–4 months after maturity, while the others were released gradually over a 7–8 month period. The temporal pattern of fruit dispersal varied with habitat but not with amount of precipitation during summer. The pattern of fruit deposition on the soil surface was affected by neighbouring plants, wind velocity, wind direction and topography. In both habitats, >90% of the fruits were deposited beside large and small clusters of plants, mainly Ephedra przewalskii. To facilitate plant community development, we suggest that E. przewalskii should be planted (as a wind break) together with Z. xanthoxylon when native pioneer species are used for restoration of cold desert shrublands.

Seed heteromorphism is the formation of different seed morphs from the same individual. Two seed morphs have been preliminarily observed in Leptocereus scopulophilus. One morph shows an apparent natural scarification of its coat. Herein we describe the seeds, taking into account shape, coat integrity, surface, dimensions, mass and the position of germination cracks. We defined two seed morphs using the integrity of the spermoderma: fragmented seed coats (FSC) and complete seed coats (CSC). We also evaluated minimum germination time, germination rate and germinability. The seed morphs did not differ significantly in traits; however, regular striations along the cuticle of the periclinal walls were more visible in the FSC compared with the CSC. Both seed morphs displayed anticlinal cell boundaries in the border region that are channelled and straight in the dorsal-ventral region but difficult to define in the lateral region. We found four morphological variations in different positions where the radicle or cotyledons emerge and variations in cuticle thickness in different regions of the seed that could determine the formation of cracks during germination. All germination variants occurred in both seed morphs, albeit in different proportions. Germination was higher and faster for the FSC compared with the CSC. These germination differences could be related to a thinner cuticle in the FSC and the punctual release of its spermoderma, which facilitates a quick imbibition of the embryo and the breaking of the seed coat. Our results indicate that differences in germination parameters between the two seed morphs relate to differences in the percentage of dormant seeds, which favour the temporal expansion of germination and reduce competition between siblings. To propagate the species for conservation purposes, we recommend using FSC, while CSC may be used to establish a seed collection ex situ.

In recent years germination experiments have become more and more complex. Typically, they are replicated in time as independent runs and at each time point they involve hierarchical, often factorial experimental designs, which are now commonly analysed by means of linear mixed models. However, in order to characterize germination in response to time elapsed, specific event-time models are needed and mixed model extensions of these models are not readily available, neither in theory nor in practice. As a practical workaround we propose a two-step approach that combines and weighs together results from event-time models fitted separately to data from each germination test by means of meta-analytic random effects models. We show that this approach provides a more appropriate appreciation of the sources of variation in hierarchically structured germination experiments as both between- and within-experiment variation may be recovered from the data.