A role for brassinosteroids in germination in Arabidopsis

This paper presents evidence that plant brassinosteroid (BR) hormones play a role in promoting germination. It has long been recognized that seed dormancy and germination are regulated by the plant hormones abscisic acid (ABA) and gibberellin (GA). These two hormones act antagonistically with each other. ABA induces seed dormancy in maturing embryos and inhibits germination of seeds. GA breaks seed dormancy and promotes germination. Severe mutations in GA biosynthetic genes in Arabidopsis, such as ga1-3, result in a requirement for GA application to germinate. Whereas previous work has shown that BRs play a critical role in controlling cell elongation, cell division, and skotomorphogenesis, no germination phenotypes have been reported in BR mutants. We show that BR rescues the germination phenotype of severe GA biosynthetic mutants and of the GA-insensitive mutant sleepy1. This result shows that BR stimulates germination and raises the possibility that BR is needed for normal germination. If true, we would expect to detect a germination phenotype in BR mutants. We found that BR mutants exhibit a germination phenotype in the presence of ABA. Germination of both the BR biosynthetic mutant det2-1 and the BR-insensitive mutant bri1-1 is more strongly inhibited by ABA than is germination of wild type. Thus, the BR signal is needed to overcome inhibition of germination by ABA. Taken together, these results point to a role for BRs in stimulating germination.     

Thermoinhibition uncovers a role for strigolactones in Arabidopsis seed germination

Strigolactones are host factors that stimulate seed germination of parasitic plant species such as Striga and Orobanche. This hormone is also important in shoot branching architecture and photomorphogenic development. Strigolactone biosynthetic and signaling mutants in model systems, unlike parasitic plants, only show seed germination phenotypes under limited growth condition. To understand the roles of strigolactones in seed germination, it is necessary to develop a tractable experimental system using model plants such as Arabidopsis. Here, we report that thermoinhibition, which involves exposing seeds to high temperatures, uncovers a clear role for strigolactones in promoting Arabidopsis seed germination. Both strigolactone biosynthetic and signaling mutants showed increased sensitivity to seed thermoinhibition. The synthetic strigolactone GR24 rescued germination of thermoinbibited biosynthetic mutant seeds but not a signaling mutant. Hormone analysis revealed that strigolactones alleviate thermoinhibition by modulating levels of the two plant hormones, GA and ABA. We also showed that GR24 was able to counteract secondary dormancy in Arabidopsis ecotype Columbia (Col) and Cape Verde island (Cvi). Systematic hormone analysis of germinating Striga helmonthica seeds suggested a common mechanism between the parasitic and non-parasitic seeds with respect to how hormones regulate germination. Thus, our simple assay system using Arabidopsis thermoinhibition allows comparisons to determine similarities and differences between parasitic plants and model experimental systems for the use of strigolactones.     

Seed germination of GA-insensitive sleepy1 mutants does not require RGL2 protein disappearance in Arabidopsis

We explore the roles of gibberellin (GA) signaling genes SLEEPY1 (SLY1) and RGA-LIKE2 (RGL2) in regulation of seed germination in Arabidopsis thaliana, a plant in which the hormone GA is required for seed germination. Seed germination failure in the GA biosynthesis mutant ga1-3 is rescued by GA and by mutations in the DELLA gene RGL2, suggesting that RGL2 represses seed germination. RGL2 protein disappears before wild-type seed germination, consistent with the model that GA stimulates germination by causing the SCF(SLY1) E3 ubiquitin ligase complex to trigger ubiquitination and destruction of RGL2. Unlike ga1-3, the GA-insensitive sly1 mutants show variable seed dormancy. Seed lots with high seed dormancy after-ripened slowly, with stronger alleles requiring more time. We expected that if RGL2 negatively controls seed germination, sly1 mutant seeds that germinate well should accumulate lower RGL2 levels than those failing to germinate. Surprisingly, RGL2 accumulated at high levels even in after-ripened sly1 mutant seeds with 100% germination, suggesting that RGL2 disappearance is not a prerequisite for seed germination in the sly1 background. Without GA, several GA-induced genes show increased accumulation in sly1 seeds compared with ga1-3. It is possible that the RGL2 repressor of seed germination is inactivated by after-ripening of sly1 mutant seeds.     

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。     

The DELLA domain of GA INSENSITIVE mediates the interaction with the GA INSENSITIVE DWARF1A gibberellin receptor of Arabidopsis

Gibberellic acid (GA) promotes seed germination, elongation growth, and flowering time in plants. GA responses are repressed by DELLA proteins, which contain an N-terminal DELLA domain essential for GA-dependent proteasomal degradation of DELLA repressors. Mutations of or within the DELLA domain of DELLA repressors have been described for species including Arabidopsis thaliana, wheat (Triticum aestivum), maize (Zea mays), and barley (Hordeum vulgare), and we show that these mutations confer GA insensitivity when introduced into the Arabidopsis GA INSENSITIVE (GAI) DELLA repressor. We also demonstrate that Arabidopsis mutants lacking the three GA INSENSITIVE DWARF1 (GID1) GA receptor genes are GA insensitive with respect to GA-promoted growth responses, GA-promoted DELLA repressor degradation, and GA-regulated gene expression. Our genetic interaction studies indicate that GAI and its close homolog REPRESSOR OF ga1-3 are the major growth repressors in a GA receptor mutant background. We further demonstrate that the GA insensitivity of the GAI DELLA domain mutants is explained in all cases by the inability of the mutant proteins to interact with the GID1A GA receptor. Since we found that the GAI DELLA domain alone can mediate GA-dependent GID1A interactions, we propose that the DELLA domain functions as a receiver domain for activated GA receptors.     

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。     

Temperature controls seed germination and dormancy in the European woodland herbaceous perennial Erythronium dens-canis (Liliaceae)

We examined the germination ecology and the temperature requirements for germination of Erythronium dens-canis, under both outdoor and laboratory conditions. E. dens-canis is a spring flowering woodland geophyte widely distributed across Europe. Germination phenology, including embryo development and radicle and cotyledon emergence, were investigated in a natural population growing in Northern Italy. Immediately after harvest, seeds of E. dens-canis were either sown on agar in the laboratory under simulated seasonal temperatures or placed in nylon mesh sachets and buried in the wild. Embryos, undifferentiated at the time of seed dispersal, grew during summer and autumn conditions in the laboratory and in the wild, culminating in radicle emergence in winter when temperatures fell to ≈ 5 °C. Emergence of cotyledons did not occur immediately after radicle emergence, but was delayed until the end of winter. Laboratory experiments showed that temperature is the main factor controlling dormancy and germination, with seeds becoming non-dormant only when given warmth, followed by cold stratification. Unlike seeds of E. dens-canis that germinate in winter, in other Erythronium species radicle emergence occurs in autumn, while in some it is delayed until seeds are transferred from winter to spring conditions. Our results suggest that there is genetic and environmental control of the expression of seed dormancy amongst Erythronium species, which is related to local climate.     

Molecular mechanisms and hormonal regulation underpinning morphological dormancy: a case study using Apium graveolens (Apiaceae)

Underdeveloped (small) embryos embedded in abundant endosperm tissue, and thus having morphological dormancy (MD) or morphophysiological dormancy (MPD), are considered to be the ancestral state in seed dormancy evolution. This trait is retained in the Apiaceae family, which provides excellent model systems for investigating the underpinning mechanisms. We investigated Apium graveolens (celery) MD by combined innovative imaging and embryo growth assays with the quantification of hormone metabolism, as well as the analysis of hormone and cell-wall related gene expression. The integrated experimental results demonstrated that embryo growth occurred inside imbibed celery fruits in association with endosperm degradation, and that a critical embryo size was required for radicle emergence. The regulation of these processes depends on gene expression leading to gibberellin and indole-3-acetic acid (IAA) production by the embryo and on crosstalk between the fruit compartments. ABA degradation associated with distinct spatiotemporal patterns in ABA sensitivity control embryo growth, endosperm breakdown and radicle emergence. This complex interaction between gibberellins, IAA and ABA metabolism, and changes in the tissue-specific sensitivities to these hormones is distinct from non-MD seeds. We conclude that the embryo growth to reach the critical size and the associated endosperm breakdown inside MD fruits constitute a unique germination programme.     

Some reflections on the relationship between endogenous hormones and light-mediated seed dormancy

The relationships between phytochrome and endogenous hormones in the light-mediated control of seed dormancy are discussed. It is concluded that gibberellins are primarily involved in post-dormancy metabolic processes leading to embryo growth and radicle emergence, such as food reserve mobilisation and endosperm softening. Evidence is considered that germination inhibitors, particularly abscisic acid, are involved in the establishment and maintenance of primary dormancy. The role of cytokinins not fully elucidated but there is considerable evidence to suggest that phytochrome control may involve cytokinin effects on transmembrane ion fluxes. In terms of hormonal control, phytochrome mediated dormancy is a complex phenomenon. There is a need for molecular studies of processes controlled by phytochrome, GAs, CKs and ABA during dormancy and germination to unravel the complexities of the dormancy mechanisms. Such studies would be facilitated by the availability of CK-deficient mutants of classical light-sensitive species.     

赤霉素作用机理的分子基础与调控模式研究进展

赤霉素(gibberellins或gibberellic acid, GA)作为植物生长的必需激素之一, 调控植物生长发育的各个方面, 如: 种子萌发, 下胚轴的伸长, 叶片的生长和植物开花时间等。近年来随着植物功能基因组学的进一步发展, 有关赤霉素生物合成及其调控, 赤霉素信号转导途径, 以及赤霉素与其他激素和环境因子的互作等领域的研究取得了较大的进展。本文综述了赤霉素生物合成的生物学途径及其调控研究; GA信号转导通道的研究进展, 特别是DELLA蛋白阻遏植物生长发育的分子机理和GA解除阻遏作用(derepress)的分子模型; GA受体研究的新进展; 探讨GA与其它激素之间的相互作用, 以及植物在应答环境过程中的作用。     

珍稀濒危植物珙桐种子休眠萌发过程中内源激素的变化

珙桐是我国特有珍稀濒危植物,休眠期长且具二次休眠现象。将处于休眠萌发过程中的珙桐种子依据胚根长度划分为4个阶段,利用高效液相色谱(HPLC)测定各阶段种子及其内果皮中ABA(脱落酸)、GA(赤霉素)、KT(细胞分裂素)、IAA(3-吲哚乙酸)4种内源激素含量,分析其比值动态变化,并与幼苗阶段进行比较。结果显示:未破壳种子的内果皮中内源激素含量以ABA最高,其次是GA、IAA、KT,随着种子破壳后四种激素含量显著降低。除ABA外,种子中GA、IAA和KT含量随着胚根的伸长逐渐升高,但仍低于幼苗阶段。此外,随着胚根伸长,种子中GA/ABA、IAA/ABA、KT/ABA比值逐渐增大,其中以GA/ABA的变化最显著。因此,珙桐种子的休眠和萌发可能主要受ABA和GA的平衡和拮抗来调控。     

Proteolysis-independent downregulation of DELLA repression in Arabidopsis by the gibberellin receptor GIBBERELLIN INSENSITIVE DWARF1

This article presents evidence that DELLA repression of gibberellin (GA) signaling is relieved both by proteolysis-dependent and -independent pathways in Arabidopsis thaliana. DELLA proteins are negative regulators of GA responses, including seed germination, stem elongation, and fertility. GA stimulates GA responses by causing DELLA repressor degradation via the ubiquitin-proteasome pathway. DELLA degradation requires GA biosynthesis, three functionally redundant GA receptors GIBBERELLIN INSENSITIVE DWARF1 (GID1a, b, and c), and the SLEEPY1 (SLY1) F-box subunit of an SCF E3 ubiquitin ligase. The sly1 mutants accumulate more DELLA proteins but display less severe dwarf and germination phenotypes than the GA biosynthesis mutant ga1-3 or the gid1abc triple mutant. Interestingly, GID1 overexpression rescued the sly1 dwarf and infertility phenotypes without decreasing the accumulation of the DELLA protein REPRESSOR OF ga1-3. GID1 rescue of sly1 mutants was dependent on the level of GID1 protein, GA, and the presence of a functional DELLA motif. Since DELLA shows increasing interaction with GID1 with increasing GA levels, it appears that GA-bound GID1 can block DELLA repressor activity by direct protein-protein interaction with the DELLA domain. Thus, a SLY1-independent mechanism for GA signaling may function without DELLA degradation.     

Gibberellin requirement for Arabidopsis seed germination is determined both by testa characteristics and embryonic abscisic acid

The mechanisms imposing a gibberellin (GA) requirement to promote the germination of dormant and non-dormant Arabidopsis seeds were analyzed using the GA-deficient mutant ga1, several seed coat pigmentation and structure mutants, and the abscisic acid (ABA)-deficient mutant aba1. Testa mutants, which exhibit reduced seed dormancy, were not resistant to GA biosynthesis inhibitors such as tetcyclacis and paclobutrazol, contrarily to what was found before for other non-dormant mutants in Arabidopsis. However, testa mutants were more sensitive to exogenous GAs than the wild-types in the presence of the inhibitors or when transferred to a GA-deficient background. The germination capacity of the ga1-1 mutant could be integrally restored, without the help of exogenous GAs, by removing the envelopes or by transferring the mutation to a tt background (tt4 and ttg1). The double mutants still required light and chilling for dormancy breaking, which may indicate that both agents can have an effect independently of GA biosynthesis. The ABA biosynthesis inhibitor norflurazon was partially efficient in releasing the dormancy of wild-type and mutant seeds. These results suggest that GAs are required to overcome the germination constraints imposed both by the seed coat and ABA-related embryo dormancy.     

血皮槭种子休眠机制研究

利用抑制物生物测定法和酸蚀技术研究了血皮槭种子休眠的原因。血皮槭种子吸水是一个非常缓慢的过程,在140 h以后种子含水量才能达到68%左右。酸蚀处理种子3 h,虽然没有加快种子的吸水速率,但能较好得使果皮变薄,也不影响种子的生活力。种子的各部位（果皮、种皮、子叶、胚根）均含有抑制物质,对小白菜种子的发芽率及胚根生长有很强的抑制作用,子叶各种处理水浸提液的抑制作用最强,果皮和种皮次之。血皮槭种子休眠主要由种壳机械障碍和种胚生理休眠两重因素导致,因此如何克服致密果壳而使激素能接触生理休眠的种胚是打破其种子休眠的关键技术。     

DELLA Proteins and GA Signalling in <Emphasis Type="Italic">Arabidopsis</Emphasis>

Gibberellin (GA) is a classical plant hormone involved in many aspects of plant growth and development. A family of five homologs called the DELLA proteins, comprised of GAI, RGA, RGL1, RGL2 and RGL3, were recently found to act as critical GA signal mediators in Arabidopsis. Reports have shown that GAI and RGA are coupled together to repress stem elongation growth whereas RGL2 is a major negative regulator of seed germination. GA down-regulates DELLA proteins through protein degradation likely via the proteasome pathway. The conserved and functionally important DELLA domain is responsible for protein stability in response to GA.     

光信号与激素调控种子休眠和萌发研究进展

休眠是种子植物在长期进化过程中产生的适应性性状, 通过抑制种子在不适宜的环境中萌发进而保证植物能够在逆境中生存。此外, 休眠有助于种子的长距离运输和扩散, 因此休眠对种子延续和物种保存具有重要意义。种子由休眠向萌发的发育转变不仅关系到物种的繁衍, 而且对保证农业生产中作物的产量和品质也具有重要作用。种子的休眠和萌发受到内源激素和外源光信号的共同调控。其中, 外源光信号主要通过调控内源ABA和GA的生物合成及信号转导进而调控种子休眠和萌发。该文系统综述了外源光信号和内源激素调控种子休眠和萌发的作用通路以及两类信号通路之间的交互作用, 旨在为农业生产中利用光和激素调控种子休眠与萌发提供参考。