The ABSCISIC ACID-INSENSITIVE 3 (ABI3) gene is expressed during vegetative quiescence processes in Arabidopsis

The ABSCISIC ACID-INSENSITIVE 3 ( ABI3 ) gene of Arabidopsis thaliana (L.) Heynh is known to play an important role during seed maturation and dormancy. Here, we present evidence suggesting an additional role for ABI3 during vegetative quiescence processes. During growth in the dark, ABI3 is expressed in the apex of the seedlings after cell division is arrested. The 2S seed storage protein gene, a target gene of ABI3 in seeds, is also induced in the arrested apex under similar darkness conditions. In addition, β -glucuronidase expression under the control of the ABI3 promoter is abolished by treatments that provoke leaf development in the dark [sucrose and abscisic acid (ABA) biosynthesis inhibitors] and induced by treatments that prevent leaf development (darkness and ABA). Furthermore, ABI3 expression is absent in apices of dark-grown de-etiolated ( det 1 ) and abi3 mutants, both known to develop leaves or leaf primordia in the dark. The fact that the expression of the ABI3 gene is only observed in a fraction of the analysed plants suggests that ABI3 is probably only one of the components of a molecular network underlying quiescence. In addition to the expression of ABI3 in apices of dark-grown seedlings, the ABI3 promoter confers expression in other vegetative organs as well, such as the stipules and the abscission zones of the siliques. In conclusion, apart from its role in seed development, ABI3 might have additional functions.     

ABA-insensitive (ABI) 4 and ABI5 synergistically regulate DGAT1 expression in Arabidopsis seedlings under stress

Triacylglycerol (TAG) accumulation is essential for seed maturation in plants. Diacylglycerol acyltransferase 1 (DGAT1) is the rate-limiting enzyme in TAG biosynthesis. In this study, we show that TAG accumulation in Arabidopsis seedlings is correlated with environmental stress, and both ABI4 and ABI5 play important roles in regulating DGAT1 expression. Tobacco transient assays revealed the synergistic effect of ABI4 with ABI5 in regulating DGAT1 expression. Taken together, our findings indicate ABI5 is an important accessory factor with ABI4 in the activation of DGAT1 in Arabidopsis seedlings under stress.     

PYR/PYL/RCAR family members are major in‐vivo ABI1 protein phosphatase 2C‐interacting proteins in Arabidopsis

Abscisic acid (ABA) mediates resistance to abiotic stress and controls developmental processes in plants. The group‐A PP2Cs, of which ABI1 is the prototypical member, are protein phosphatases that play critical roles as negative regulators very early in ABA signal transduction. Because redundancy is thought to limit the genetic dissection of early ABA signalling, to identify redundant and early ABA signalling proteins, we pursued a proteomics approach. We generated YFP‐tagged ABI1 Arabidopsis expression lines and identified in vivo ABI1‐interacting proteins by mass‐spectrometric analyses of ABI1 complexes. Known ABA signalling components were isolated including SnRK2 protein kinases. We confirm previous studies in yeast and now show that ABI1 interacts with the ABA‐signalling kinases OST1, SnRK2.2 and SnRK2.3 in plants. Interestingly, the most robust in planta ABI1‐interacting proteins in all LC‐MS/MS experiments were nine of the 14 PYR/PYL/RCAR proteins, which were recently reported as ABA‐binding signal transduction proteins, providing evidence for in vivo PYR/PYL/RCAR interactions with ABI1 in Arabidopsis. ABI1–PYR1 interaction was stimulated within 5 min of ABA treatment in Arabidopsis. Interestingly, in contrast, PYR1 and SnRK2.3 co‐immunoprecipitated equally well in the presence and absence of ABA. To investigate the biological relevance of the PYR/PYLs, we analysed pyr1/pyl1/pyl2/pyl4 quadruple mutant plants and found strong insensitivities in ABA‐induced stomatal closure and ABA‐inhibition of stomatal opening. These findings demonstrate that ABI1 can interact with several PYR/PYL/RCAR family members in Arabidopsis, that PYR1–ABI1 interaction is rapidly stimulated by ABA in Arabidopsis and indicate new SnRK2 kinase‐PYR/PYL/RCAR interactions in an emerging model for PYR/PYL/RCAR‐mediated ABA signalling.     

A BIN2-GLK1 Signaling Module Integrates Brassinosteroid and Light Signaling to Repress Chloroplast Development in the Dark

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New cross talk between ROS,ABA and auxin controlling seed maturation and germination unraveled in APX6 deficient Arabidopsis seeds

Successful execution of germination program greatly depends on the seeds’ oxidative homeostasis. We recently identified new roles for the H₂O₂-reducing enzyme ascorbate peroxidase 6 (APX6) in germination control and seeds’ stress tolerance. APX6 replaces APX1 as the dominant APX in dry seeds, and its loss-of-function results in reduced germination due to over accumulation of ROS and oxidative damage. Metabolic analyses in dry apx6 seeds, revealed altered homeostasis of primary metabolites including accumulation of TCA cycle metabolites, ABA and auxin, supporting a novel role for APX6 in regulating cellular metabolism. Increased sensitivity of apx6 mutants to ABA or IAA in germination assays indicated impaired perception of these signals. Relative suppression of ABI3 and ABI5 expression, and induction of ABI4, suggested the activation of a signaling route inhibiting germination in apx6 seeds that is independent of ABI3. Here we provide additional evidence linking ABI4 with ABA- and auxin-controlled inhibition of germination and suggest a hypothetical model for the role of APX6 in the regulation of the crosstalk between these hormones and ROS.     

Partial complementation of embryo defective mutations: a general strategy to elucidate gene function

The EMB 506 gene has been characterised as essential for embryo development. To provide insights into the role of EMB 506, which is hidden by the embryo defective phenotype, the ABI3 promoter was fused to the EMB 506 cDNA. The expression of such a transgene should provide sufficient protein during embryogenesis to ensure normal embryo development in homozygous emb 506 seeds. We show that homozygous emb 506 seedlings, partially complemented with the ABI3::EMB 506 transgene, can be obtained. Most of the rescued emb 506 plants are able to flower and to set normal seeds, but show mild to severe depigmentation of rosette leaves and/or inflorescences. This effect on chloroplast development indicated a putative chloroplast localisation of the EMB 506 protein, which was demonstrated by GFP-protein fusion. However, EMB 506 cannot be considered as a chloroplast housekeeping protein only, since EMB 506 is not present in all photosynthetic tissues. This study demonstrates the power of this simple strategy, which could be widely applied to other emb mutants and which may reveal similar or additional roles for EMB genes at vegetative stages of the life cycle.     

花生ABI3同源基因的定位分析

ABSC ISIC AC ID-INSITIVE3(AB I3)、LEAFY COTYLEDON2(LEC2)和FUSCA3(FUS3)转录因子在种子发育过程中发挥着重要的调控作用。采用Northern杂交技术,用拟南芥AB I3保守的B3结构域部分序列作为探针分别与花生根、茎、叶、子叶RNA进行了杂交,同时也对花生根、茎、叶、子叶(含胚)组织切片进行了原位杂交,结果均显示只有在花生的子叶和胚中有杂交信号出现,表明花生中可能存在AB I3、FUS3和LEC2的同源基因,且它们只分布在花生的子叶和胚中。     

异源过表达齿肋赤藓ScABI3基因改变拟南芥气孔表型并提高抗旱性

ABI3是ABA信号通路中关键的转录调控因子, 参与种子休眠、质体发育及苔藓耐干等重要生理过程, 在植物抗逆中发挥关键作用。以荒漠耐干苔藓——齿肋赤藓(Syntrichia caninervis)为材料, 克隆了抗逆基因ScABI3并获得3个独立的pCAMBIA1301-ScABI3转基因拟南芥(Arabidopsis thaliana)纯合株系。结果表明, 转基因拟南芥叶片气孔孔径增大, 单位面积气孔数量减少, 植株水分利用效率提高; 在干旱处理14天后转基因拟南芥植株存活率显著高于野生型, 离体叶片失水率显著低于野生型。进一步研究发现, ScABI3转基因拟南芥通过提高自身活性氧(ROS)清除能力增强植株抗旱性。研究结果可为开发利用荒漠植物基因资源培育抗逆作物品种奠定基础。