Unlike sporadic daytime heat spikes, a consistent increase in night‐time temperatures can potentially derail the genetic gains being achieved. Ten winter wheat genotypes were exposed to six different night‐time temperatures (15–27°C) during flowering and grain‐filling stages in controlled environment chambers. We identified the night‐time temperature of 23oC as the critical threshold beyond which a consistent decline in yields and quality was observed. Confocal laser scanning micrographs of central endosperm, bran, and germ tissue displayed differential accumulation of protein, lipid, and starch with increasing night‐time temperatures. KS07077M‐1 recorded a decrease in starch and an increase in protein and lipid in central endosperm with increasing night‐time temperatures, whereas the same was significantly lower in the tolerant SY Monument. Expression analysis of genes encoding 21 enzymes (including isoforms) involved in grain–starch metabolism in developing grains revealed a high night‐time temperature (HNT)‐induced reduction in transcript levels of adenosine diphosphate glucose pyrophosphorylase small subunit involved in starch synthesis and a ≥2‐fold increase in starch degrading enzymes isoamylase III, alpha‐, and beta‐amylase. The identified critical threshold, grain compositional changes, and the key enzymes in grain starch metabolism that lead to poor starch accumulation in grains establish the foundational knowledge for enhancing HNT tolerance in wheat.

Sorghum is often exposed to suboptimal low temperature stress under field conditions, particularly at the seedling establishment stage. Enhancing chilling tolerance will facilitate earlier planting and so minimize the negative impacts of other stresses experienced at later growth stages. Genome-wide association mapping was performed on a sorghum association panel grown under control (30/20 °C; day/night) and chilling (20/10 °C) conditions. Genomic regions on chromosome 7, controlling the emergence index and seedling (root and shoot) vigor, were associated with increased chilling tolerance but they did not co-localize with undesirable tannin content quantitative trait loci (QTLs). Shoot and root samples from highly contrasting haplotype pairs expressing differential responses to chilling stress were used to identify candidate genes. Three candidate genes (an alpha/beta hydrolase domain protein, a DnaJ/Hsp40 motif-containing protein, and a YTH domain-containing RNA-binding protein) were expressed at significantly higher levels under chilling stress in the tolerant haplotype compared with the sensitive haplotype and BTx623. Moreover, two CBF/DREB1A transcription factors on chromosome 2 showed a divergent response to chilling in the contrasting haplotypes. These studies identify haplotype differences on chromosome 7 that modulate chilling tolerance by either regulating CBF or feeding back into this signaling pathway. We have identified new candidate genes that will be useful markers in ongoing efforts to develop tannin-free chilling-tolerant sorghum hybrids.

BackgroundMaintenance of seed viability is an important factor for seedling vigour and plant establishment. Lipid peroxidation mediated reactive carbonyl compounds (RCC’s) and non-enzymatic modifications of proteins through Maillard and Amadori products reduce seed viability and seedling vigour.ResultsIn this study, the relevance of RCCs on genotypic variation in rice seed viability and overexpression of an aldo-ketoreductase (AKR1) enzyme that detoxify cytotoxic compounds to improve seed viability and vigour was studied. Physiological and biochemical approaches were integrated to quantify the variation in seed viability and seedling vigour in rice genotypes after exposing to ageing treatment. AKR1 was overexpressed in a susceptible rice genotype and tobacco to study the relevance of reduced RCC’s on seed viability and seedling vigour. The glycation and lipid peroxidation compounds accumulated after accelerated ageing treatments in rice genotypes. The accumulation of malondialdehyde, methyl glyoxal, Maillard and Amadori products affected the seed viability and germination as they showed a significant negative relationship. The transgenic rice and tobacco seeds expressing AKR1 showed lower levels of cytotoxic compounds and glycation products that resulted in improved seed viability and seedling vigour in rice and tobacco.ConclusionsThe study demonstrates that, reactive cytotoxic compounds affect the seed viability during storage. Detoxification of reactive cytotoxic compounds by Aldo-keto reductases is one of the mechanisms to improve the seed longevity during storage.Electronic supplementary materialThe online version of this article (doi:10.1186/s12284-017-0148-3) contains supplementary material, which is available to authorized users.

Using existing protocols, RNA extracted from seeds rich in starch often results in poor quality RNA, making it inappropriate for downstream applications. Though some methods are proposed for extracting RNA from plant tissue rich in starch and other polysaccharides, they invariably yield less and poor quality RNA. In order to obtain high yield and quality RNA from seeds and other plant tissues including roots a modified SDS-LiCl method was compared with existing methods, including TRIZOL kit (Invitrogen), Plant RNeasy mini kit (Qiagen), Furtado (2014) method, and CTAB-LiCl method. Modifications in the extraction buffer and solutions used for RNA precipitation resulted in a robust method for extracting RNA in seeds and roots, where extracting quality RNA is challenging. The modified SDS-LiCl method revealed intense RNA bands through gel electrophoresis and a nanodrop spectrophotometer detected ratios of ≥ 2 and 1.8 for A260/A230 and A260/A280, respectively. The absence of starch co-precipitation during RNA extraction resulted in enhanced yield and quality of RNA with RIN values of 7–9, quantified using a bioanalyzer. The high-quality RNA obtained was demonstrated to be suitable for downstream applications, such as cDNA synthesis, gene amplification, and RT-qPCR. The method was also effective in extracting RNA from seeds of other cereals including field-grown sorghum and corn. The modified SDS-LiCl method is a robust and highly reproducible RNA extraction method for plant tissues rich in starch and other secondary metabolites. The modified SDS-LiCl method successfully extracted high yield and quality RNA from mature, developing, and germinated seeds, leaves, and roots exposed to different abiotic stresses.

SummaryIn recent years, concerns about the use of glyphosate‐resistant crops have increased because of glyphosate residual levels in plants and development of herbicide‐resistant weeds. In spite of identifying glyphosate‐detoxifying genes from microorganisms, the plant mechanism to detoxify glyphosate has not been studied. We characterized an aldo‐keto reductase gene from Pseudomonas (PsAKR1) and rice (OsAKR1) and showed, by docking studies, both PsAKR1 and OsAKR1 can efficiently bind to glyphosate. Silencing AKR1 homologues in rice and Nicotiana benthamiana or mutation of AKR1 in yeast and Arabidopsis showed increased sensitivity to glyphosate. External application of AKR proteins rescued glyphosate‐mediated cucumber seedling growth inhibition. Regeneration of tobacco transgenic lines expressing PsAKR1 or OsAKRI on glyphosate suggests that AKR can be used as selectable marker to develop transgenic crops. PsAKR1‐ or OsAKRI‐expressing tobacco and rice transgenic plants showed improved tolerance to glyphosate with reduced accumulation of shikimic acid without affecting the normal photosynthetic rates. These results suggested that AKR1 when overexpressed detoxifies glyphosate in planta.