拟南芥突变体种子休眠与萌发的研究进展

种子的休眠和萌发是一个复杂的过程,至今尚未能清楚阐明其调控机制。目前已从拟南芥突变体中鉴定了一些与种子萌发和休眠相关的基因,有助于阐明种子休眠和萌发的分子机制。本文综述了拟南芥突变体种子休眠与萌发方面的研究进展。赤霉素是促进种子萌发的主要因素之一,RGL、SPY、GCR、SLY和GAR等基因的表达参与赤霉素对种子萌发的调控。脱落酸与种子休眠有关,ABI1、ABI2、ABI3、ABI4、ABI5、FUS3、LEC、MARD和CIPK等基因参与了脱落酸的调控过程。对3类乙烯反应的突变体（ein、etr和ctr）以及油菜素内酯突变体（det和bri）的研究表明乙烯和油菜素内酯是通过拮抗脱落酸而促进种子萌发的。光对种子萌发的调节,是通过具有Ser／Thr蛋白激酶活性的光敏色素PhyA、PhyB、PhyC、PhyD和PhyE,以磷酸化／去磷酸化方式调节其它与萌发相关基因的表达。含氮化合物对种子萌发的促进,可能是以一种依赖一氧化氮的方式解除种子休眠。     

Seed after-ripening is a discrete developmental pathway associated with specific gene networks in Arabidopsis

After-ripening (AR) is a time and environment regulated process occurring in the dry seed, which determines the germination potential of seeds. Both metabolism and perception of the phytohormone abscisic acid (ABA) are important in the initiation and maintenance of dormancy. However, molecular mechanisms that regulate the capacity for dormancy or germination through AR are unknown. To understand the relationship between ABA and AR, we analysed genome expression in Arabidopsis thaliana mutants defective in seed ABA synthesis (aba1-1) or perception (abi1-1). Even though imbibed mutant seeds showed no dormancy, they exhibited changes in global gene expression resulting from dry AR that were comparable with changes occurring in wild-type (WT) seeds. Core gene sets were identified that were positively or negatively regulated by dry seed storage. Each set included a gene encoding repression or activation of ABA function (LPP2 and ABA1, respectively), thereby suggesting a mechanism through which dry AR may modulate subsequent germination potential in WT seeds. Application of exogenous ABA to after-ripened WT seeds did not reimpose characteristics of freshly harvested seeds on imbibed seed gene expression patterns. It was shown that secondary dormancy states reinstate AR status-specific gene expression patterns. A model is presented that separates the action of ABA in seed dormancy from AR and dry storage regulated gene expression. These results have major implications for the study of genetic mechanisms altered in seeds as a result of crop domestication into agriculture, and for seed behaviour during dormancy cycling in natural ecosystems.     

The Time Required for Dormancy Release in Arabidopsis Is Determined by DELAY OF GERMINATION1 Protein Levels in Freshly Harvested Seeds

Seed dormancy controls the start of a plant's life cycle by preventing germination of a viable seed in an unfavorable season. Freshly harvested seeds usually show a high level of dormancy, which is gradually released during dry storage (after-ripening). Abscisic acid (ABA) has been identified as an essential factor for the induction of dormancy, whereas gibberellins (GAs) are required for germination. The molecular mechanisms controlling seed dormancy are not well understood. DELAY OF GERMINATION1 (DOG1) was recently identified as a major regulator of dormancy in Arabidopsis thaliana. Here, we show that the DOG1 protein accumulates during seed maturation and remains stable throughout seed storage and imbibition. The levels of DOG1 protein in freshly harvested seeds highly correlate with dormancy. The DOG1 protein becomes modified during after-ripening, and its levels in stored seeds do not correlate with germination potential. Although ABA levels in dog1 mutants are reduced and GA levels enhanced, we show that DOG1 does not regulate dormancy primarily via changes in hormone levels. We propose that DOG1 protein abundance in freshly harvested seeds acts as a timer for seed dormancy release, which functions largely independent from ABA.     

Seed dormancy and ABA metabolism in Arabidopsis and barley: the role of ABA 8'-hydroxylase

We have investigated the relationship between seed dormancy and abscisic acid (ABA) metabolism in the monocot barley and the dicot Arabidopsis. Whether dormant (D) or non-dormant (ND), dry seed of Arabidopsis and embryos of dry barley grains all had similarly high levels of ABA. ABA levels decreased rapidly upon imbibition, although they fell further in ND than in D. Gene expression profiles were determined in Arabidopsis for key ABA biosynthetic [the 9-cis epoxycarotenoid dioxygenasegene family] and ABA catabolic [the ABA 8'-hydroxylase gene family (CYP707A)] genes. Of these, only the AtCYP707A2 gene was differentially expressed between D and ND seeds, being expressed to a much higher level in ND seeds. Similarly, a barley CYP707 homologue, (HvABA8'OH-1) was expressed to a much higher level in embryos from ND grains than from D grains. Consistent with this, in situ hybridization studies showed HvABA8'OH-1 mRNA expression was stronger in embryos from ND grains. Surprisingly, the signal was confined in the coleorhiza, suggesting that this tissue plays a key role in dormancy release. Constitutive expression of a CYP707A gene in transgenic Arabidopsis resulted in decreased ABA content in mature dry seeds and a much shorter after-ripening period to overcome dormancy. Conversely, mutating the CYP707A2 gene resulted in seeds that required longer after-ripening to break dormancy. Our results point to a pivotal role for the ABA 8'-hydroxylase gene in controlling dormancy and that the action of this enzyme may be confined to a particular organ as in the coleorhiza of cereals.     

拟南芥突变体种子休眠与萌发的研究进展

种子的休眠和萌发是一个复杂的过程, 至今尚未能清楚阐明其调控机制。目前已从拟南芥突变体中鉴定了一些与种子萌发和休眠相关的基因, 有助于阐明种子休眠和萌发的分子机制。本文综述了拟南芥突变体种子休眠与萌发方面的研究进展。赤霉素是促进种子萌发的主要因素之一, RGL、SPY、GCR、SLY和GAR等基因的表达参与赤霉素对种子萌发的调控。脱落酸与种子休眠有关, ABI1、ABI2、ABI3、ABI4、ABI5、FUS3、LEC、MARD和CIPK等基因参与了脱落酸的调控过程。对3类乙烯反应的突变体 (ein、etr和ctr) 以及油菜素内酯突变体 (det和bri) 的研究表明乙烯和油菜素内酯是通过拮抗脱落酸而促进种子萌发的。光对种子萌发的调节, 是通过具有Ser/Thr蛋白激酶活性的光敏色素PhyA、PhyB、 PhyC、PhyD和PhyE, 以磷酸化/去磷酸化方式调节其它与萌发相关基因的表达。含氮化合物对种子萌发的促进, 可能是以一种依赖一氧化氮的方式解除种子休眠。     

Control of seed dormancy in Nicotiana plumbaginifolia: post-imbibition abscisic acid synthesis imposes dormancy maintenance

The physiological characteristics of seed dormancy in Nicotiana plumbaginifolia Viv. are described. The level of seed dormancy is defined by the delay in seed germination (i.e the time required prior to germination) under favourable environmental conditions. A wild-type line shows a clear primary dormancy, which is suppressed by afterripening, whereas an abscisic acid (ABA)-deficient mutant shows a non-dormant phenotype. We have investigated the role of ABA and gibberellic acid (GA₃) in the control of dormancy maintenance or breakage during imbibition in suitable conditions. It was found that fluridone, a carotenoid biosynthesis inhibitor, is almost as efficient as GA₃ in breaking dormancy. Dry dormant seeds contained more ABA than dry afterripened seeds and, during early imbibition, there was an accumulation of ABA in dormant seeds, but not in afterripened seeds. In addition, fluridone and exogenous GA₃ inhibited the accumulation of ABA in imbibed dormant seeds. This reveals an important role for ABA synthesis in dormancy maintenance in imbibed seeds. Received: 31 December 1998 / Accepted: 9 July 1999     

Engineering seed dormancy by the modification of zeaxanthin epoxidase gene expression

Abscisic acid (ABA) is a plant hormone synthesized during seed development that is involved in the induction of seed dormancy. Delayed germination due to seed dormancy allows long-term seed survival in soil but is generally undesirable in crop species. Freshly harvested seeds of wild-type Nicotiana plumbaginifolia plants exhibit a clear primary dormancy that results in delayed germination, the degree of primary dormancy being influenced by environmental culture conditions of the mother plant. In contrast, seeds, obtained either from ABA-deficient mutant aba2-s1 plants directly or aba2-s1 plants grafted onto wild-type plant stocks, exhibited rapid germination under all conditions irrespective of the mother plant culture conditions. The ABA biosynthesis gene ABA2 of N. plumbaginifolia, encoding zeaxanthin epoxidase, was placed under the control of the constitutive 35S promoter. Transgenic plants overexpressing ABA2 mRNA exhibited delayed germination and increased ABA levels in mature seeds. Expression of an antisense ABA2 mRNA, however, resulted in rapid seed germination and in a reduction of ABA abundance in transgenic seeds. It appears possible, therefore, that seed dormancy can be controlled in this Nicotiana model species by the manipulation of ABA levels.     

Molecular mechanisms underlying the entrance in secondary dormancy of Arabidopsis seeds

As seasons change, dormant seeds cycle through dormant states until the environmental conditions are favourable for seedling establishment. Dormancy cycle is widespread in the plant kingdom allowing the seeds to display primary and secondary dormancy. Several reports in the last decade have focused on understanding the molecular mechanisms of primary dormancy, but our knowledge regarding secondary dormancy is limited. Here, we studied secondary dormancy induced in Arabidopsis thaliana by incubating seeds at 25 °C in darkness for 4 d. By physiological, pharmacological, expression and genetics approaches, we demonstrate that (1) the entrance in secondary dormancy involves changes in the content and sensitivity to GA, but the content and sensitivity to ABA do not change, albeit ABA is required; (2) RGL2 promotes the entrance in secondary dormancy through ABI5 action; and (3) multivariate analysis with 18 geographical and environmental parameters of accession collection place suggests that temperature is an important variable influencing the induction of secondary dormancy in nature.     

Functional analysis in Arabidopsis of FsPTP1, a tyrosine phosphatase from beechnuts,reveals its role as a negative regulator of ABA signaling and seed dormancy and suggests its involvement in ethylene signaling modulation

By means of an RT-PCR approach we isolated a specific tyrosine phosphatase (FsPTP1) induced by abscisic acid (ABA) and correlated with seed dormancy in Fagus sylvatica seeds. To provide genetic evidence of FsPTP1 function in seed dormancy and ABA signal transduction pathway, we overexpressed this gene in Cape Verde Island ecotype of Arabidopsis thaliana, which shows the deepest degree of seed dormancy among Arabidopsis accessions. As a result, 35S:FsPTP1 transgenic seeds showed a reduced dormancy and insensitivity to ABA and osmotic stress conditions accompanied by a reduction in the level of expression of RAB18 and RD29, well-known ABA-responsive genes. Taken together, all these data are consistent with a role of this tyrosine phosphatase as a negative regulator of ABA signaling. In addition, phenotypes of FsPTP1 transgenic plants resemble those observed in ethylene constitutive mutants, accompanied by an increase in the level of expression of a key gene involved in ethylene signaling such as EIN2. All the data presented along the paper suggest that the effect of tyrosine phosphatases in ABA action during the transition from seed dormancy to germination may be through modulation of ethylene signaling.     

生长素调控种子的休眠与萌发

植物种子的休眠与萌发,是植物生长发育过程中的关键阶段,也是生命科学领域的研究热点。种子从休眠向萌发的转换是极为复杂的生物学过程,由外界环境因子、体内激素含量及信号传导和若干关键基因协同调控。大量研究表明,植物激素脱落酸(Abscisic acid, ABA)和赤霉素(Gibberellin acid, GA)是调控种子休眠水平,决定种子从休眠转向萌发的主要内源因子。ABA与GA在含量和信号传导两个层次上的精确平衡,确保了植物种子能以休眠状态在逆境中存活,并在适宜的时间启动萌发程序。生长素(Auxin)是经典植物激素之一,其对向性生长和组织分化等生物学过程的调控已有大量研究。但最近有研究证实,生长素对种子休眠有正向调控作用,这表明生长素是继ABA之后的第二个促进种子休眠的植物激素。本文在回顾生长素的发现历程、阐释生长素体内合成途径及信号传导通路的基础上,重点综述了生长素通过与ABA的协同作用调控种子休眠的分子机制,并对未来的研究热点进行了讨论和展望。     

Antagonistic effects of abscisic acid and gibberellic acid on the breaking of dormancy of Fagus sylvatica seeds

Study of the factors involved in the dormancy of Fagus sylvatica seeds shows that such dormancy is due partly to the seed coats and partly to endogenous factors. Seed coat removal accelerates both the release from dormancy and the effects of the other treatments that abolish it. The dormancy of these seeds is eliminated by cold treatment at 4°C over a period longer than 8 weeks, and exogenous application of abscisic acid (ABA) reverses the effects of low temperature, the seeds remaining in an ungerminated state. Additionally, ABA reduces protein synthesis but slightly increases RNA synthesis, which suggests its involvement in the synthesis of RNAs related to this process. In vitro translation of the RNAs isolated from these seeds shows that ABA delays the disappearance of at least 2 polypeptides (of ca 22 and 24 kDa), which are abundant in dormant seeds and under conditions that prevent the release from dormancy, but which disappear under treatments that abolish it. Exogenous application of gibberellic acid (GA₃) proved to be efficient in breaking the dormancy of these seeds and in substituting for cold treatment as well as in antagonizing the effects of ABA on the synthesis of both DNA and proteins. GA₃ also accelerates the disappearance of the two polypeptides abundant in dormant seeds and in ABA-treated seeds. These findings suggest that both ABA and GA₃ could be involved in the regulation of nucleic acid and protein metabolism during dormancy, acting antagonistically in these processes and, specifically, in the regulation of the synthesis of the two proteins that appear to play a role in the maintenance of dormancy in these seeds.