拟南芥AtR8 lncRNA对盐胁迫响应及其对种子萌发的调节作用

长链非编码RNA (lncRNA)是一类长度大于200个核苷酸且不编码蛋白质的非编码RNA, 主要由RNA聚合酶II转录生成, 大量存在于生物体内并具有多种生物学功能。AtR8 lncRNA是拟南芥(Arabidopsis thaliana)中RNA聚合酶III转录的长链非编码RNA。前期研究发现, 水杨酸(SA)处理诱导萌发种子中AtR8 lncRNA的表达, AtR8 lncRNA缺失抑制SA胁迫下的种子萌发。进一步研究发现, AtR8 lncRNA转录区域内存在保守的盐胁迫响应元件(TCTTCTTCTTTA); NaCl处理抑制萌发种子中AtR8 lncRNA的表达; 与野生型相比, 高浓度NaCl处理明显抑制了atr8 (AtR8 lncRNA部分缺失型拟南芥)种子萌发。研究结果表明, AtR8 lncRNA在拟南芥种子萌发期的盐胁迫中起重要作用。     

温度和盐分胁迫对新疆鼠尾草种子萌发的影响

利用控制实验研究了温度和盐分胁迫对新疆鼠尾草种子萌发的影响.结果显示:新疆鼠尾草种子萌发的最适温度为25℃.盐胁迫对新疆鼠尾草种子的萌发均有明显的抑制作用,随着盐浓度的增加,萌发率和萌发速率下降.在适宜萌发温度下,新疆鼠尾草耐盐极限值0.2 mmol/L.     

Arabinan Metabolism during Seed Development and Germination in Arabidopsis

Arabinans are found in the pectic network of many cell walls, where, along with galactan, they are present as side chains of Rhamnogalacturonan I. Whilst arabinans have been reported to be abundant polymers in the cell walls of seeds from a range of plant species, their proposed role as a storage reserve has not been thoroughly investigated. In the cell walls of Arabidopsis seeds, arabinose accounts for approximately 40% of the monosaccharide composition of non- cellulosic polysaccharides of embryos. Arabinose levels decline to -15% during seedling establishment, indicating that cell wall arabinans may be mobilized during germination. Immunolocalization of arabinan in embryos, seeds, and seedlings reveals that arabinans accumulate in developing and mature embryos, but disappear during germination and seedling establishment. Experiments using 14C-arabinose show that it is readily incorporated and metabolized in growing seedlings, indicating an active catabolic pathway for this sugar. We found that depleting arabinans in seeds using a fungal arabinanase causes delayed seedling growth, lending support to the hypothesis that these polymers may help fuel early seedling growth.     

The Arabidopsis LARP1s are Involved in Regulation of Seed Germination

Members of La-related protein (LARP) 1 family spread widely in various species, and they are involved in regulating many important biological processes in mammal, yeast, and fruit fly. However, functional research of LARP1s in plants is limited so far. In Arabidopsis, there are three members in LARP1 family, LARP1a, 1b, and 1c. Here, we found that the mutation of LARP1 genes delayed seed germination, implying that LARP1 proteins might be positive factors of seed germination in Arabidopsis. Moreover, the larp1 mutants showed more sensitive to abscisic acid (ABA) and paclobutrazol (PAC), as larp1 mutants displayed low rate of germination in medium contained ABA or PAC. Temporal and spatial expression analyses revealed that LARP1s were more abundant in seeds, especially in imbibed seeds. Subcellular localization analysis revealed that all LARP1 proteins could localize to the P-bodies, suggesting that LARP1s might play a role in RNA processes. Taken together, our results unravel new conserved functions of LARP1s in the regulation of Arabidopsis seed germination.

     

盐胁迫对4种短命植物种子萌发及植株生长的影响

以新疆北部4种十字花科短命植物为材料,研究不同浓度NaCl胁迫下其种子萌发及植株生长特性,以明确它们的耐盐性强弱。结果显示:(1)随着NaCl浓度的增加,4种植物种子萌发率、萌发指数和相对萌发率均不同程度下降,而相对盐害率则逐渐升高,并以绵果荠的耐盐性最好,其在300 mmol.L-1NaCl胁迫下仍有22.22%的萌发率。(2)4种植物的生长在50~150 mmol.L-1NaCl胁迫下没有受到明显抑制,在200 mmol.L-1浓度下受不同程度的抑制,而在300 mmol.L-1NaCl胁迫下抑制作用更严重,其中绵果荠在各浓度处理下长势均最好,能够在300 mmol.L-1NaCl胁迫下存活10 d。(3)在盐胁迫条件下,4种植物细胞质膜透性不同程度增加,而叶片相对含水量和光合色素含量均不同程度降低,并以绵果荠各项指标的变化幅度最小。可见,4种新疆短命植物种子萌发和植株生长均受到盐胁迫的伤害,但它们的耐盐阈值明显不同,并以绵果荠的耐盐性最强。     

温度和水分及盐分胁迫对银沙槐种子萌发的影响

利用控制实验研究了温度、湿度、干旱和盐分胁迫等生态因子对银沙槐种子萌发的影响,以探索银沙槐种子对各种生态因子的适应性。结果显示:(1)银沙槐种子在20℃、25℃恒温和15℃/25℃、20℃/30℃、10℃/20℃变温环境中的发芽率较高且无显著差异,其在20℃恒温、15℃/25℃、20℃/30℃变温条件下的发芽指数较高,但差异不显著。(2)土壤含水量在1%～5%之间,各水分处理间种子发芽率差异显著(P<0.05),而在5%～25%间种子发芽率变化不显著。(3)盐胁迫和水分胁迫对银沙槐种子的萌发均有明显的抑制作用,可显著降低种子萌发率(P<0.05);种子发芽指数和活力指数均随渗透势和NaCl浓度增大而显著减小(P<0.05);恢复萌发率随渗透势和NaCl浓度的增加而显著增加(P<0.05)。研究发现,银沙槐种子萌发最适温度为20℃恒温和15℃/25℃变温,最适土壤含水量为10%～25%;种子萌发对盐分和干旱胁迫表现出不同程度的耐受性,萌发过程中主导抑制因素为渗透胁迫,离子毒害作用甚微;银沙槐种子休眠机制和萌发特征表现出它对生境的良好适应性。     

Ethylene receptor ETHYLENE RECEPTOR1 domain requirements for ethylene responses in Arabidopsis seedlings

Ethylene influences many processes in Arabidopsis (Arabidopsis thaliana) through the action of five receptor isoforms. We used high-resolution, time-lapse imaging of dark-grown Arabidopsis seedlings to better understand the roles of each isoform in the regulation of growth in air, ethylene-stimulated nutations, and growth recovery after ethylene removal. We found that ETHYLENE RECEPTOR1 (ETR1) is both necessary and sufficient for nutations. Transgene constructs in which the ETR1 promoter was used to drive expression of cDNAs for each of the five receptor isoforms were transferred into etr1-6;etr2-3;ein4-4 triple loss-of-function mutants that have constitutive growth inhibition in air, fail to nutate in ethylene, and take longer to recover a normal growth rate when ethylene is removed. The patterns of rescue show that ETR1, ETR2, and ETHYLENE INSENSITIVE4 (EIN4) have the prominent roles in rapid growth recovery after removal of ethylene whereas ETR1 was the sole isoform that rescued nutations. ETR1 histidine kinase activity and phosphotransfer through the receiver domain are not required to rescue nutations. However, REVERSION TO SENSITIVITY1 modulates ethylene-stimulated nutations but does not modulate the rate of growth recovery after ethylene removal. Several chimeric receptor transgene constructs where domains of EIN4 were swapped into ETR1 were also introduced into the triple mutant. The pattern of phenotype rescue by the chimeric receptors used in this study supports a model where a receptor with a receiver domain is required for normal growth recovery and that nutations specifically require the full-length ETR1 receptor.     

The Copper Transporter RAN1 Is Essential for Biogenesis of Ethylene Receptors in Arabidopsis

Plants utilize ethylene as a hormone to regulate multiple developmental processes and to coordinate responses to biotic and abiotic stress. In Arabidopsis thaliana, a small family of five receptor proteins typified by ETR1 mediates ethylene perception. Our previous work suggested that copper ions likely play a role in ethylene binding. An independent study indicated that the ran1 mutants, which display ethylene-like responses to the ethylene antagonist trans-cyclooctene, have mutations in the RAN1 copper-transporting P-type ATPase, once again linking copper ions to the ethylene-response pathway. The results presented herein indicate that genetically engineered Saccharomyces cerevisiae expressing ETR1 but lacking the RAN1 homolog Ccc2p (Δccc2) lacks ethylene-binding activity. Ethylene-binding activity was restored when copper ions were added to the Δccc2 mutants, showing that it is the delivery of copper that is important. Additionally, transformation of the Δccc2 mutant yeast with RAN1 rescued ethylene-binding activity. Analysis of plants carrying loss-of-function mutations in ran1 showed that they lacked ethylene-binding activity, whereas seedlings carrying weak alleles of ran1 had normal ethylene-binding activity but were hypersensitive to copper-chelating agents. Altogether, the results show an essential role for RAN1 in the biogenesis of the ethylene receptors and copper homeostasis in Arabidopsis seedlings. Furthermore, the results indicate cross-talk between the ethylene-response pathway and copper homeostasis in Arabidopsis seedling development.     

Low Pressure and Ethylene in Lettuce Seed Germination

The extent and manner of ethylene involvement in germination of lettuce (Lactuca Sativa L. cv. Mesa 659) seed at a moderate temperature (20°C) were investigated. Inhibition of germination at low pressure of 150 mmHg in an oxygen flow-through system was alleviated to a marked extent by ethylene. Carbon dioxide was ineffective by itself but caused further alleviation of inhibition in presence of ethylene and oxygen. Other seed treatments which partially alleviated the inhibition caused by low pressure included soaking in 10μM of fusicoccin and a prior treatment with acetone. Of the two ethylene adsorbents used, Purafil was more effective in inhibiting germination in a closed container. Although the ethylene biosynthesis inhibitor, 8-hydroxyquinoline (1.0 mM). showed no effect on ethylene production, it markedly inhibited germination and the effect was partially reversed by ethylene and GA₃. An ethoxy analog of rhizobitoxine, on the other hand, had little or no effect on germination but strongly inhibited the ethylene production. Although no causal relation of ethylene to germination was established, the evidence presented here implicates ethylene, together with other gases, in the regulation of germination.     

盐胁迫对不同品种小白菜种子萌发特性的影响

采用5个小白菜(Brassica chinensis)品种(‘七宝青’、‘夏冬青’、‘四月慢’、‘南京中杆’和‘605’),用100 mmol/L NaCl对种子发芽作盐胁迫处理,从发芽势、发芽率、发芽指数、盐害指数、胚长和胚鲜重等指标比较了5个小白菜品种的抗盐性。结果表明,‘七宝青’种子的耐盐性优于其它品种。     

Knockout of Multiple Arabidopsis Cation/H+ Exchangers Suggests Isoform-Specific Roles in Metal Stress Response,Germination and Seed Mineral Nutrition

Cation/H⁺ exchangers encoded by CAX genes play an important role in the vacuolar accumulation of metals including Ca²⁺ and Mn²⁺. Arabidopsis thaliana CAX1 and CAX3 have been previously shown to differ phylogenetically from CAX2 but the physiological roles of these different transporters are still unclear. To examine the functions and the potential of redundancy between these three cation transporters, cax1/cax2 and cax2/cax3 double knockout mutants were generated and compared with wild type and cax single knockouts. These double mutants had equivalent metal stress responses to single cax mutants. Both cax1 and cax1/cax2 had increased tolerance to Mg stress, while cax2 and cax2/cax3 both had increased sensitivity to Mn stress. The cax1/cax2 and cax2/cax3 mutants did not exhibit the deleterious developmental phenotypes previously seen with the cax1/cax3 mutant. However, these new double mutants did show alterations in seed germination, specifically a delay in germination time. These alterations correlated with changes in nutrient content within the seeds of the mutants, particularly the cax1/cax2 mutant which had significantly higher seed content of Ca and Mn. This study indicates that the presence of these Arabidopsis CAX transporters is important for normal germination and infers a role for CAX proteins in metal homeostasis within the seed.