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Transcriptome and physiological analysis of rapeseed tolerance to post-flowering temperature increase

ABSTRACT: This study investigates the transcriptome and physiological responses of rapeseed to post-flowering temperature increases, providing valuable insights into the molecular mechanisms underlying rapeseed tolerance to heat stress. Two rapeseed genotypes, Lumen and Solar, were assessed under control and heat stress conditions in field experiments conducted in Valdivia, Chile. Results showed that seed yield and seed number were negatively affected by heat stress, with genotype-specific responses. Lumen exhibited a 9.3% average seed yield reduction, while Solar showed a 28.7% reduction. RNA-seq analysis of siliques and seeds revealed tissue-specific responses to heat stress, with siliques being more sensitive to temperature stress. Hierarchical clustering analysis identified distinct gene clusters reflecting different aspects of heat stress adaptation in siliques, with a role for protein folding in maintaining silique development and seed quality under high temperature conditions. In seeds, three distinct patterns of heat-responsive gene expression were observed, with genes involved in protein folding and response to heat showing genotype-specific expression. Gene coexpression network analysis revealed major modules for rapeseed yield and quality, as well as the trade-off between seed number and seed weight. Overall, this study contributes to understanding the molecular mechanisms underlying rapeseed tolerance to heat stress and can inform crop improvement strategies targeting yield optimization under changing environmental conditions.

ORGANISM(S): Brassica napus

PROVIDER: GSE241954 | GEO | 2023/09/02

REPOSITORIES: GEO

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Project description:Effects of heat priming applied to the first generation on tolerance of the successive generation to post-anthesis high temperature stress were investigated. Compared with the progeny of non-heat primed plants (NH), the progeny of heat-primed plants (PH) presented higher grain yield, leaf photosynthesis and activities of antioxidant enzymes and lower cell membrane damage under high temperature stress. In the transcriptome profile, 1430 probes showed obvious difference in expression between PH and NH. These genes were those of signal transduction, transcription, energy, defense, and protein destination and storage, respectively. Pot experiments (25 cm in diameter and 22 cm in depth) were conducted at the Experimental Station of Nanjing Agricultural University, Nanjing, China (32Ë30Ê¹N and 118Ë42Ê¹E). During the first generation, wheat plants were divided into two groups: one group were not primed (N) while the other were heat-primed (P) at pre-anthesis (at a day/night temperature of 32/28ËC for two days at the seven-leaf stage and the nine-leaf stage) and post-anthesis (34/30ËC for seven days at the 10th day after anthesis). At maturity, seeds from both groups were harvested. Seedlings (the progeny generation) from each group seed were further divided two sub-groups: one was subjected to high temperature stress at a day/night temperature of 34/30ËC, while the other was set at 26/22ËC for six days from the 10th day after anthesis (DAA). During both priming and high-temperature stress, plants under different treatments were moved into sepatarate growth chambers at preset temperatures. Thereafter, four treatments were established: NC, progeny of non-primed plants without post-anthesis high temperature stress; NH, progeny of non-primed plants with post-anthesis high temperature stress; PC, progeny of primed plants without post-anthesis high temperature stress; and PH, progeny of primed plants with post-anthesis high temperature stress.

Project description:There are 16 organ samples (dry seeds, 24H imbibed seeds, 48H imbibed seeds, juvenile rosette, adult rosette, senescence leaves, cauline leaves, stems, young buds, mature flower buds, flowers, young siliques, mature siliques and old siliques) with triplicates. There are 17 samples of different environmental samples (0 h white, 1 h white, 6 h white, 24 h white, dark, blue, far-red and red lights, control, cold 2h, cold 6h, hot 2h, hot 6h, NaCl 2h, NaCl 6h, dry 2h and dry 6h) with triplicates. For 12 organs, three lines of plants were grown in soil under the long-day conditions of 16 hours light at 22℃. Samples were collected from each of juvenile rosette leaves, adult rosette leaves, cauline leaves, stems, young flower buds (sepals were closed and petals are not visible), mature flower buds (petals are visible), flowers, young siliques filled with white seeds, mature siliques filled with green seeds, old yellowing siliques (2-month-old) and senescence rosettes. Root tissues were collected from 2-week-old plants growing on 1% agar plates containing Murashige and Skoog (MS). Callus were collected from cultured CIM (callus induction medium) containing B5 medium supplemented with 20 g liter–1 glucose, 0.5 mg liter–1 2,4-D, and 0.1 mg liter–1 kinetin. Dry seed was defined to be after-ripened seeds for 4 weeks. Imbibed seeds are collected from seeds imbibed on the moistened paper for 24 and 48 hours under the continuous light at 22℃. In light irradiations, wild type (WT) seeds were plated on MS plates. For the time course experiment of white light, the plates were placed at 4°C in darkness for at least 2 days prior to receiving 6 h red light irradiation to synchronize germination. Then, plates were placed in the dark for 3 days and then exposed to white fluorescent light (100 μmol m–2 s–1) for 1, 6 and 24 h. For monochromatic light experiment, WT seedlings were grown in continuous red light, blue light, far-red light, or darkness for 6 days after red light irradiation. The light intensity used was 0.17 μmol m–2 s–1 for red light, 1.16 μmol m–2 s–1 for blue light and 0.53 μmol m–2 s–1 for far-red light. All abiotic stress samplings were collected from plants grown for 2 weeks on MS plates under 2 and 6 hours of abiotic stress conditions at the continuous light at 22℃. For drought stress treatment, plants were transferred and dehydrated onto the dried filter paper in the covered plastic dishes. For heat and cold stress treatments, the covered plastic dishes with plants were transferred directly from 22 to 37 and 2℃ respectively. For salinity stress treatment, plants were transferred onto the filter paper moistened with MS plates including 200mM NaCl.

Project description:To better undersand the effects of drought stress on wheat developing seeds, the transcription profile of early developing wheat seeds under control and drought stress conditions were comparatively analyzed by using the Affymetrix wheat geneChip. Drought stress is a major yield-limiting factor for wheat. Wheat yields are particularly sensitive to drought stress during reproductive development. Early seed development stage is an important determinant of seed size, one of the yield components. We specifically examined the impact of drought stress imposed during postzygotic early seed development in wheat. We imposed a short-term drought stress on plants with day-old seeds and observed that even a short-duration drought stress significantly reduced the size of developing seeds as well as mature seeds. Drought stress delayed the developmental transition from syncytial to cellularized stage of endosperm. Coincident with reduced seed size and delayed endosperm development, a subset of genes associated with cytoskeleton organization was misregulated in developing seeds under drought-stressed. Several genes linked to hormone pathways were also differentially regulated in response to drought stress in early seeds. Notably, drought stress strongly repressed the expression of wheat storage protein genes such as gliadins, glutenins and avenins as early as 3 days after pollination. Our results provide new insights on how some of the early seed developmental events are impacted by water stress, and the underlying molecular pathways that can possibly impact both grain size and quality in wheat. Winter wheat cultivar Redland, PI 502907 (Triticum aestivum L.) was used for this study. Seedlings were vernalized at 4°C for 6 weeks and then transplanted to a one gallon pot of soil-sand mixture (3:1, v/v) and grown in a growth chamber under the following conditions: relative humidity, 50–70%; 16-h light/8-h dark photoperiod; 21°C daytime temperature and 18°C nights. Plants were watered regularly twice daily at the rate of 100ml/ per pot. Because, wheat has an asynchronous fertilization pattern for ovlues in the inflorescence, each floret needs to be specifically marked for timing the fertilization and stress induction. After spikes developed, unfertilized ovules were monitored and observed for the fertilization process. Closed wheat spikes with anthers outside were marked as fertilized. Drought stress was imposed 24h after the fertilization (HAF). Drought stress treatment was initiated by discontinuing watering on the drought treatment plants while control plants were regularly watered twice daily. Stress treatment was applied at 48 HAF and relieved at 96 HAF. The microarray study focuses on 24 HAF to 72 HAF in control and drought stress conditions. We started to impose drought stress 24HAF.

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Transcriptome and physiological analysis of rapeseed tolerance to post-flowering temperature increase

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