低等植物蓝光受体信号传导研究

拟南芥含有5个已分离的蓝光受体和至少1个未鉴定的蓝光/紫外光-A受体.隐花色素(CRY1、CRY2和CRY3) 调节植物的形态建成、开花和生物节律性,而向光素 (PHOT1和PHOT2) 调节植物的向光性、叶绿体运动和气孔开放.黄素可以吸收蓝光和紫外光-A,是CRY和PHOT蓝光受体的生色团.对这些光受体的结构和作用模式已了解很多.苔藓植物小立碗藓中含有2个已分离的隐花色素(CRY1a和CRY1b),负责调节侧枝形成和调控生长素反应;有4个向光素(PHOTA1,PHOTA2,PHOTB1,PHOTB2) 调节叶绿体的运动.苔藓细胞内蓝光/紫外光-A引发的信号转导有Ca2+参与.     

植物蓝光受体研究进展

植物蓝光调节的反应主要有向光性、抑制幼茎伸长、叶绿体迁移、刺激气孔张开和调节基因表达等。蓝光反应的有效波长是蓝光和近紫外光(320—400am),故蓝光受体也叫蓝光／近紫外光受体。植物蓝光受体研究近年来取得较大进展。以拟南芥为例,已得到确认的受体至少有隐花色素(CRY1、2)和向光蛋白(phototropin)两大类。转基因拟南芥对蓝光、紫外光和绿光敏感,并发现CRY1是一个可溶性蛋白。CRY2编码一个核蛋白,蓝光在转录水平对该蛋白进行调节,它的作用是增加拟南芥对蓝光的敏感性。CRY1和CRY2共同介导了拟南芥植物的向光性。隐花色素的蛋白与辅基之间以非共价键连接,可以吸收蓝光和近紫外光。CRY1和CRY2蛋白之间,尤其是N端相似性很高。向光蛋白目前只发现PHOT1和PHO12两种,向光蛋白作为丝／苏氨酸激酶蓝光受体含有两个光氧化结构域(LOV)并参与了植物向光性叶绿体运动、气孔开放等。     

向光素调节植物向光性及其与光敏色素/隐花色素的相互关系

蓝光受体向光素(PHOT1/PHOT2)调节蓝光诱导的植物运动反应, 包括植物向光性、叶绿体运动、气孔运动和叶片伸展等。其中, 向光素介导的植物向光性能够促使植物弯向光源, 确保其以最佳取向捕获光源, 优化光合作用。光敏色素和隐花色素作为光受体也参与植物的向光性调节。该文综述了向光素介导的拟南芥(Arabidopsis thaliana)下胚轴向光弯曲信号转导及其与光敏色素、隐花色素协同作用的分子机制, 以期为改造植物光捕获能力及提高光利用效率提供理论基础。     

植物下胚轴向光弯曲机制研究进展

植物向光弯曲生长主要是由于其向光和背光面生长素的不对称分布引起。近年来研究发现,在不同强度的蓝光单侧照射下,植物可能存在不同的向光弯曲调节机制。目前,关于向光素PHOT1介导弱蓝光引起的下胚轴弯曲研究较为详细,即PHOT1感受蓝光后,与其下游的信号蛋白NPH3、RPT2和PKS1相互作用,调控生长素运输蛋白的活性及定位,诱导生长素的不对称分布引起向光弯曲。PHOT1和PHOT2以功能冗余方式调节强蓝光引起的植物下胚轴向光弯曲,NPH3可能作为共享调节因子,引发不同的信号转导通路实现功能互补。此外,其他光受体、激素、蛋白激酶、蛋白磷酸酶以及Ca2＋也参与了植物向光弯曲的调节。本文就近年来有关植物下胚轴向光弯曲信号组分及可能的网络关系进行总结,并对该研究领域存在的问题及今后可能的研究方向进行展望。     

植物向光素受体与信号转导机制研究进展

向光素(phototropin,PHOT)是继光敏色素、隐花色素之后分离的植物蓝光受体。PHOT介导蓝光诱导的向光反应,叶绿体运动,气孔开放、叶片伸展及叶片定位等生理反应。近年来关于PHOT受体介导这些生理反应的分子机制探讨愈来愈受研究者的广泛关注。主要从拟南芥PHOT结构及信号转导方面的研究进展进行综述。     

拟南芥蓝光受体CRY2的功能及其介导的光形态建成

植物具备一套复杂的由两种蓝光受体和多种信号转导下游组分组成的蓝光感应系统,通过感受光照强度、光的方向和光周期,调节自身对蓝光的应答.蓝光反应的有效波长是蓝光和近紫外光(320～400nm),故蓝光受体也叫蓝光/近紫外光受体.CRY2(Cryptochromes,CRY)是一个核蛋白,在转录水平受蓝光的调节,它的作用是增加拟南芥对蓝光的敏感性.植物蓝光调节的反应主要有向光性、抑制幼茎伸长、叶绿体迁移、刺激气孔张开和调节基因表达等.对植物蓝光反应突变体分子生物学研究进展进行了综述,对蓝光受体及信号转导下游组分在植物发育中的作用及蓝光诱发植物作出反应的分子机制进行了探讨.     

隐花色素蛋白光依赖磁感应机制研究进展

隐花色素(Cryptochrome,CRY)蛋白是一种对蓝光敏感的蛋白,在植物中主要调节生长及发育,是果蝇等昆虫光信号的接收者,在哺乳动物中扮演调节生物钟的角色。CRY蛋白作为磁光信号感受器受到广泛关注,其结构中的保守的色氨酸三联体与辅因子黄素腺嘌呤二核苷酸(flavin adenine dinucleotide,FAD)间自由基电子对是CRY蛋白具有光依赖的磁感应功能的关键因素。该文对CRY蛋白的分类及结构特征、光依赖磁感应机制的研究进展进行了综述,并对目前CRY蛋白研究中存在的问题及未来研究方向进行了展望。     

拟南芥ZTL/FKF1/LKP2蛋白家族功能研究进展

植物通过各类受体来感知外界环境的改变从而调节自身的生长和发育情况。在拟南芥中,植物主要通过隐花色素(Cryptochromes)和向光素(Phototropins)感知蓝光。同时ZEITLUPE (ZTL),FLAVIN-BINDING KELCH REPEAT F-box1 (FKF1)和LOV KELCH PROTEIN2 (LKP2)蛋白家族也作为蓝光受体参与调控植物生长发育过程。因其特殊的蛋白结构组成,在植物的光周期开花、节律性和光形态建成等方面发挥了重要的调控作用。近来,ZTL/FKF1/LKP2蛋白家族被发现参与植物逆境胁迫响应。本文归纳了ZTL/FKF1/LKP2的生物学功能研究进展,并对其作用机制进行了总结与讨论。     

PHOT2介导拟南芥下胚轴向光弯曲调节子的筛选与鉴定

向光素(PHOT1和PHOT2)功能冗余调节单侧强蓝光诱导的拟南芥(Arabidopsis thaliana)黄化苗下胚轴向光弯曲表现功能冗余,限制了人们对PHOT2信号转导机制的深入研究。通过化学诱变剂甲基磺酸乙酯(EMS)诱变拟南芥phot1突变体,避开PHOT1基因的干扰,寻找PHOT2下游信号分子。研究结果表明,已成功筛选到1株遗传稳定的下胚轴向蓝光不弯曲突变体。遗传分析结果显示,该突变体可能是PHOT2下游信号分子突变,将其命名为p2sa1(phototropin2 signaling associated1)。用100μmol·m–2·s–1强蓝光单侧照射,phot1p2sa1下胚轴向光弯曲缺失,呈现phot1phot2双突变的表型,然而phot1p2sa1在强蓝光下叶绿体避光正常,明显不同于phot1phot2。实验证实P2SA1可能位于PHOT2的下游,参与调节PHOT2介导的拟南芥下胚轴向光弯曲反应。     

高梁CRY2基因的克隆及其表达分析

从高粱(Sorghum bicolor L. var.R1ll)幼苗中提取总RNA,利用RT-PCR和cDNA的3'末端的快速扩增方法(3'RACE),第一次克隆了高粱隐花色素2基因(CRY2)的cDNA序列.该序列包括了一个完整的开放阅读框,编码大小为690个氨基酸残基的蛋白质,与水稻、番茄和拟南芥CRY2蛋白质的同源性分别为8 7%、5 7%和45.5%.高粱CRY2基因组DNA含有3个内含子和4个外显子.RT-PCR检测结果表明,高粱CRY2基因在根、茎和叶中都有转录.Western blotting结果显示CRY2蛋白在根、茎和叶中表达,并在黑暗中积累,蓝光下降解.高粱CRY2可能在蓝光诱导的幼苗去黄化反应中起作用.     

拟南芥NPH3/RPT2-Like (NRL)家族蛋白在向光素信号转导通路中的作用研究进展

向光素PHOT1和PHOT2感受蓝光刺激后发生自磷酸化激活, 调节植物气孔开放、叶绿体运动、叶片伸展和定位以及向光性(包括根的负向光性和下胚轴的向光性)等多种适应性反应。拟南芥(Arabidopsis thaliana) NRL (NPH3/RPT2-Like)家族成员在向光素介导的信号途径中发挥重要作用, 其中NPH3特异调控下胚轴的向光性以及叶片的伸展与定位, RPT2参与调节植物向光性、叶片的伸展与定位以及叶绿体聚光反应等。NCH1是新发现的NRL家族成员, 与RPT2以功能冗余的方式调节叶绿体的聚光反应, 但不调节避光反应。该文主要综述了NRL蛋白家族成员在向光素介导蓝光信号通路中的作用, 并展望了未来的研究方向, 旨在为全面揭示NRL家族成员的功能提供线索。     

Light-dependent, dark-promoted interaction between Arabidopsis cryptochrome 1 and phytochrome B proteins

Plant photoreceptors transduce environmental light cues to downstream signaling pathways, regulating a wide array of processes during growth and development. Two major plant photoreceptors with critical roles in photomorphogenesis are phytochrome B (phyB), a red/far-red absorbing photoreceptor, and cryptochrome 1 (CRY1), a UV-A/blue photoreceptor. Despite substantial genetic evidence for cross-talk between phyB and CRY1 pathways, a direct interaction between these proteins has not been observed. Here, we report that Arabidopsis phyB interacts directly with CRY1 in a light-dependent interaction. Surprisingly, the interaction is light-dissociated; CRY1 interacts specifically with the dark/far-red (Pr) state of phyB, but not with the red light-activated (Pfr) or the chromophore unconjugated form of the enzyme. The interaction is also regulated by light activation of CRY1; phyB Pr interacts only with the unstimulated form of CRY1 but not with the photostimulated protein. Further studies reveal that a small domain extending from the photolyase homology region (PHR) of CRY1 regulates the specificity of the interaction with different conformational states of phyB. We hypothesize that in plants, the phyB/CRY1 interaction may mediate cross-talk between the red/far-red- and blue/UV-sensing pathways, enabling fine-tuning of light responses to different spectral inputs.     

Phenotypic characterization of a photomorphogenic mutant

Light is arguably the most important abiotic factor controlling plant growth and development throughout their life cycle. Plants have evolved sophisticated light-sensing mechanisms to monitor fluctuations in light quality, intensity, direction and periodicity (day length). In Arabidopsis, three families of photoreceptors have been identified by molecular genetic studies. The UV-A/blue light receptors cryptochromes and the red/far-red receptors phytochromes control an overlapping set of responses including photoperiodic flowering induction and de-etiolation. Phototropins are the primary photoreceptors for a set of specific responses to UV-A/blue light such as phototropism, chloroplast movement and stomatal opening. Mutants affecting a photoreceptor have a characteristic phenotype. It is therefore possible to determine the specific developmental responses and the photoreceptor pathway(s) affected in a mutant by performing an appropriate set of photobiological and genetic experiments. In this paper, we outline the principal and easiest experiments that can be performed to obtain a first indication about the nature of the photobiological defect in a given mutant.     

The role of a 14-3-3 protein in stomatal opening mediated by PHOT2 in Arabidopsis

The 14-3-3 λ isoform is required for normal stomatal opening mediated by PHOT2 in Arabidopsis thaliana. Arabidopsis phototropin2 (PHOT2) interacts with the λ-isoform 14-3-3 protein both in yeast two-hybrid screening and in an in vitro pull-down assay. Further yeast two-hybrid analysis also showed that the PHOT2 C-terminal kinase domain was required for the interaction. Site-directed mutagenesis indicated that PHOT2 Ser-747 is essential for the yeast interaction. Phenotypic characterization of a loss-of-function 14-3-3 λ mutant in a phot1 mutant background showed that the 14-3-3 λ protein was necessary for normal PHOT2-mediated blue light-induced stomatal opening. PHOT2 Ser-747 was necessary for complementation of the blue light-activated stomatal response in a phot1 phot2 double mutant. The 14-3-3 λ mutant in the phot1 mutant background allowed normal phototropism and normal chloroplast accumulation and avoidance responses. It also showed normal stomatal opening mediated by PHOT1 in a phot2 mutant background. The 14-3-3 κ mutant had no effect on stomatal opening in response to blue light. Although the 14-3-3 λ mutant had no chloroplast movement phenotype, the 14-3-3 κ mutation caused a weaker avoidance response at an intermediate blue light intensity by altering the balance between the avoidance and accumulation responses. The results highlight the strict specificity of phototropin-mediated signal transduction pathways.     

New insights into resistance protein-mediated signaling against turnip crinkle virus in Arabidopsis

The Arabidopsis-turnip crinkle virus (TCV) system is one of the few tractable plant-virus systems that allow simultaneous characterization of host components required for basal- and/or resistance (R) protein-mediated defenses. Another unique feature is that hypersensitive response (HR) and resistance can be studied as two distinct phenotypes in this pathosytem. The R protein HRT confers HR to TCV but requires a recessive locus rrt to confer resistance. The pathways leading to HR and resistance are mutually exclusive. HRT interacts with EDS1, which potentiates HR to TCV and is also required for resistance signaling. HRT-mediated signaling is also dependent on the EDS1-interacting proteins PAD4 and SAG101, which form binary and ternary complexes with EDS1. HRT-mediated resistance is also dependent on light and more specifically on the blue-light photoreceptors, cryptochromes (CRY) and phototropins (PHOT). Of these, CRY2 and PHOT2 are required for the stability of HRT. HRT is degraded in a proteasome-dependent manner, which correlates with its interaction with the E3 ubiquitin ligase, COP1. Together, these results suggest that components of light signaling modulate plant defense against TCV by regulating the stability of, and signaling mediated by, the R protein HRT.     

Action spectrum for cryptochrome-dependent hypocotyl growth inhibition in Arabidopsis

Cryptochrome blue-light photoreceptors are found in both plants and animals and have been implicated in numerous developmental and circadian signaling pathways. Nevertheless, no action spectrum for a physiological response shown to be entirely under the control of cryptochrome has been reported. In this work, an action spectrum was determined in vivo for a cryptochrome-mediated high-irradiance response, the blue-light-dependent inhibition of hypocotyl elongation in Arabidopsis. Comparison of growth of wild-type, cry1cry2 cryptochrome-deficient double mutants, and cryptochrome-overexpressing seedlings demonstrated that responsivity to monochromatic light sources within the range of 390 to 530 nm results from the activity of cryptochrome with no other photoreceptor having a significant primary role at the fluence range tested. In both green- and norflurazon-treated (chlorophyll-deficient) seedlings, cryptochrome activity is fairly uniform throughout its range of maximal response (390-480 nm), with no sharply defined peak at 450 nm; however, activity at longer wavelengths was disproportionately enhanced in CRY1-overexpressing seedlings as compared with wild type. The action spectrum does not correlate well with the absorption spectra either of purified recombinant cryptochrome photoreceptor or to that of a second class of blue-light photoreceptor, phototropin (PHOT1 and PHOT2). Photoreceptor concentration as determined by western-blot analysis showed a greater stability of CRY2 protein under the monochromatic light conditions used in this study as compared with broad band blue light, suggesting a complex mechanism of photoreceptor activation. The possible role of additional photoreceptors (in particular phytochrome A) in cryptochrome responses is discussed.     

The phototropin family as photoreceptors for blue light-induced chloroplast relocation

Blue light-induced chloroplast accumulation and avoidance relocation movements are controlled by the blue light photoreceptor phototropin. The Arabidopsis thaliana genome has two phototropin genes encoding phot1 and phot2. Each of these photoreceptors contains two LOV (light oxygen and voltage) domains and a kinase domain. The LOV domains absorb blue light though an associated flavin mononucleotide chromophore, while the kinase domain is thought to be associated with signal transduction. The phototropins control not only chloroplast relocation movement, but also blue light-induced phototropic responses, leaf expansion and stomatal opening. Here I review the role of phototropin as a photoreceptor for chloroplast photorelocation movement. Electronic Publication