苔藓植物蓝光/紫外光-A 信号传导的研究

种子植物含有5个已分离的光受体和至少1个未鉴定的蓝光/紫外光-A受体。隐花色素(CRY1、CRY2和CRY3) 调节植物的生长发育,而向光蛋白(PHOT1和PHOT2) 调节植物对光的营养反应。黄素可以吸收蓝光和紫外光-A,是生色团。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生长素代谢;有4个向光蛋白(PHOTA1,PHOTA2,PHOTB1,PHOTB2) 调节叶绿体的运动。苔藓细胞内蓝光/紫外光-A刺激引发的信号转导有Ca2+参与。     

Cryptochrome 1 controls tomato development in response to blue light

Cryptochrome genes (CRY) are a novel class of plant genes encoding proteins that bear a strong resemblance to photolyases, a rare class of flavoproteins that absorb light in the blue (B) and UV-A regions of the spectrum and utilise it for photorepair of UV-damaged DNA. In Arabidopsis, both CRY1 and CRY2 are implicated in numerous blue light-dependent responses, including inhibition of hypocotyl elongation, leaf and cotyledon expansion, pigment biosynthesis, stem growth and internode elongation, control of flowering time and phototropism. No information about the in vivo function of CRY genes is available in other plant species. The tomato CRY1 gene (TCRY1) encodes a protein of 679 amino acids, which shows 78% identity and 88% similarity to Arabidopsis CRY1. In order to verify the in vivo function of TCRY1, we constructed antisense tomato plants using the C-terminal portion of the gene. Partial repression of both mRNA and protein levels was observed in one of the transformants. The progeny from this transformant showed an elongated hypocotyl under blue but not under red light. This character co-segregated with the transgene and was dependent on transgene dosage. An additional, partially elongated phenotype was observed in adult plants grown in the greenhouse under dim light and short days with no artificial illumination. This phenotype was suppressed by artificial illumination of both short and long photoperiods. The synthesis of anthocyanins under blue light was reduced in antisense seedlings. In contrast, carotenoid and chlorophyll levels and second positive phototropic curvature were essentially unaltered.     

Arabidopsis cryptochrome 1 is a soluble protein mediating blue light-dependent regulation of plant growth and development

Cryptochrome 1 (CRY1) is a flavin-type blue light receptor of Arabidopsis thaliana which mediates inhibition of hypocotyl elongation. In the work described in this report it is demonstrated that CRY1 is a soluble protein expressed in both young seedlings grown either in the dark or under light, and in different organs of adult plants. The functional role of CRY1 was further investigated using transgenic Arabidopsis plants overexpressing CRY1. It is demonstrated that overexpression of CRY1 resulted in hypersensitivity to blue, UV-A, and green light for the inhibition of hypocotyl elongation response. Transgenic plants overexpressing CRY1 also exhibited a dwarf phenotype with reduced size in almost every organ. This was in keeping with the previous observation of reciprocal alterations found in hy4 mutant plants and is consistent with a hypothesis that CRY1 mediates a light-dependent process resulting in a general inhibitory effect on plant growth. In addition, transgenic plants overexpressing CRY1 showed increased anthocyanin accumulation in response to blue, UV-A, and green light in a fluence rate-dependent manner. This increase in anthocyanin accumulation in transgenic plants was shown to be concomitant with increased blue light-induction of CHS gene expression. It is concluded that CRY1 is a photoreceptor mediating blue light-dependent regulation of gene expression in addition to its affect on plant growth.     

The role of mutants in the search for the photoreceptor for phototropism in higher plants

Early attempts to identify the chromophore of the photoreceptor for phototropism are reviewed. Carotenoids and flavins were the principal candidates, but studies with grass coleoptiles devoid of carotenoids suggest that at least in these organs carotenoids are most unlikely to play that role. The status of characterization of a gene for a putative photoreceptor protein is also reviewed. As the action spectrum for phototropism resembles the absorption spectrum of a flavoprotein, flavoproteins are attractive candidates at present, especially since the CRY1 photoreceptor in Arabidopsis thaliana that mediates blue light-dependent hypocotyl growth suppression has flavin adenine dinucleotide as one of its two chromophores. As the second chromophore appears to be pterin, pterins should not be ruled out as candidate chromophores for the photoreceptor for phototropism.     

Phototropins 1 and 2: versatile plant blue-light receptors

Blue and ultraviolet-A light regulate a wide range of responses in plants, including phototropism, chloroplast migration and stomatal opening. However, the photoreceptors for these light responses have been identified only recently. The phototropins (phot1 and phot2) represent a new class of receptor kinases that appear to be exclusive to plants. Recent genetic analysis has shown that phot1 and phot2 exhibit partially overlapping functions in mediating phototropism, chloroplast migration, and stomatal opening in Arabidopsis. Although significant progress has been made in understanding the early photochemical and biochemical events that follow phototropin excitation, the details of how this excitation activates such different responses remain to be elucidated.     

拟南芥中光信号系统中关键基因CRY1、CRY2和COP1启动子的表达模式分析

通过构建表达光信号系统关键基因CRY1、CRY2和COP1启动子与GUS融合基因的拟南芥转基因植株,并对转基因植株进行GUS组织化学染色的结果表明,CRY1、CRY2和COP1的表达模式不受光条件的调控,并且在各器官有广泛的表达。分别分析CRY1基因启动子在cop1突变体以及COP1基因启动子在cry1突变体遗传背景中表达模式的结果表明,CRY1和COP1在转录水平上不存在明显的相互调控关系。     

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。     

Manipulation of the blue light photoreceptor cryptochrome 2 in tomato affects vegetative development, flowering time, and fruit antioxidant content

Cryptochromes are blue light photoreceptors found in plants, bacteria, and animals. In Arabidopsis, cryptochrome 2 (cry2) is involved primarily in the control of flowering time and in photomorphogenesis under low-fluence light. No data on the function of cry2 are available in plants, apart from Arabidopsis (Arabidopsis thaliana). Expression of the tomato (Solanum lycopersicum) CRY2 gene was altered through a combination of transgenic overexpression and virus-induced gene silencing. Tomato CRY2 overexpressors show phenotypes similar to but distinct from their Arabidopsis counterparts (hypocotyl and internode shortening under both low- and high-fluence blue light), but also several novel ones, including a high-pigment phenotype, resulting in overproduction of anthocyanins and chlorophyll in leaves and of flavonoids and lycopene in fruits. The accumulation of lycopene in fruits is accompanied by the decreased expression of lycopene beta-cyclase genes. CRY2 overexpression causes an unexpected delay in flowering, observed under both short- and long-day conditions, and an increased outgrowth of axillary branches. Virus-induced gene silencing of CRY2 results in a reversion of leaf anthocyanin accumulation, of internode shortening, and of late flowering in CRY2-overexpressing plants, whereas in wild-type plants it causes a minor internode elongation.     

Tomato CRY1a plays a critical role in the regulation of phytohormone homeostasis,plant development,and carotenoid metabolism in fruits

Blue light photoreceptors, cryptochromes (CRYs), regulate multiple aspects of plant growth and development. However, our knowledge of CRYs is predominantly based on model plant Arabidopsis at early growth stage. In this study, we elucidated functions of CRY1a gene in mature tomato (Solanum lycopersicum) plants by using cry1a mutants and CRY1a‐overexpressing lines (OE‐CRY1a‐1 and OE‐CRY1a‐2). In comparison with wild‐type plants, cry1a mutants are relatively tall, accumulate low biomass, and bear more fruits, whereas OE‐CRY1a plants are short stature, and they not only flower lately but also bear less fruits. RNA‐seq, qRT‐PCR, and LC‐MS/MS analysis revealed that biosynthesis of gibberellin, cytokinin, and jasmonic acid was down‐regulated by CRY1a. Furthermore, DNA replication was drastically inhibited in leaves of OE‐CRY1a lines, but promoted in cry1a mutants with concomitant changes in the expression of cell cycle genes. However, CRY1a positively regulated levels of soluble sugars, phytofluene, phytoene, lycopene, and ß‐carotene in the fruits. The results indicate the important role of CRY1a in plant growth and have implications for molecular interventions of CRY1a aimed at improving agronomic traits.     

Negative phototropic response of rhizoid cells in the fern Adiantum capillus-veneris

In general, phototropic responses in land plants are induced by blue light and mediated by blue light receptor phototropins. In many cryptogam plants including the fern Adiantum capillus-veneris, however, red as well as blue light effectively induces a positive phototropic response in protonemal cells. In A. capillus-veneris, the red light effect on the tropistic response is mediated by phytochrome 3 (phy3), a chimeric photoreceptor of phytochrome and full-length phototropin. Here, we report red and blue light-induced negative phototropism in A. capillus-veneris rhizoid cells. Mutants deficient for phy3 lacked red light-induced negative phototropism, indicating that under red light, phy3 mediates negative phototropism in rhizoid cells, contrasting with its role in regulating positive phototropism in protonemal cells. Mutants for phy3 were also partially deficient in rhizoid blue light-induced negative phototropism, suggesting that phy3, in conjunction with phototropins, redundantly mediates the blue light response.     

A Spectroscopic View of Some Recent Advances in the Study of Blue Light Photoreception*

The blue to UV-A region of the spectrum, spanning the region of about 320–520 nm, strongly influences the growth and development of plants and fungi. Photomorphogenesis in plants is, to a great extent, controlled by phytochrome, but there are unique contributions of the blue region, which cannot be duplicated by any amount of red light. Phototropism is, with few exceptions, a purely blue light response. In fungi, the blue region dominates the photocontrol of growth and development, though some red light effects have been reported. Many blue light action spectra fit the definition of cryptochrome, a pigment class defined by its UV-A and blue peaks. The action spectrum, if measured to sufficient resolution, displays several minor maxima or shoulders in the blue region which call to mind the vibrational levels of carotenoids and flavins. Recent molecular genetic studies, as well as photobiological work, have shown that some cryptochromes are related to the DNA repair enzyme photolyase, while others appear genetically and spectroscopically distinct. In this review, we have applied established criteria from photobiology, in particular, comparison of action spectra with absorption spectra, to these recent results. It is apparent that photolyase homologs such as CRY1 can explain the blue light portion of the action spectrum for hypocotyl elongation, assuming participation of the oxidized flavin. In fungi, the photoreceptor question remains open. Identification of the nph1 gene in Arabidopsis may soon lead to a photoreceptor for higher plant phototropism. Also, we present a possible solution to the most recent version of the long-standing flavin-carotenoid controversy, the zeaxanthin hypothesis for higher plant phototropism. In conclusion, there appear to be at least three classes of cryptochromes.     

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。     

Stem sensitivity and ethylene involvement in phototropism of mung bean

A system is described for the examination of phototropism in the epicotyl of a dicot seedling, mung bean (Phaseolus aureus Roxb.), under conditions approximating nature, including the use of intact, nonetiolated plants exposed to elevated, continuous, white, unilateral light. It is found that in this system perception of the phototropic stimulus by the leaves alone cannot account for the curvature, and that exposure of the stem is also necessary. The phototropic response was found to be strongly altered in nonintact plants. Hypobaric treatment indicates that ethylene may participate in phototropism, possibly by acting as an inhibitor of auxin transport.     

Blue light-dependent interactions of CRY1 with GID1 and DELLA proteins regulate gibberellin signaling and photomorphogenesis in Arabidopsis

Cryptochromes are blue light photoreceptors that mediate various light responses in plants and mammals. In Arabidopsis (Arabidopsis thaliana), cryptochrome 1 (CRY1) mediates blue light-induced photomorphogenesis, which is characterized by reduced hypocotyl elongation and enhanced anthocyanin production, whereas gibberellin (GA) signaling mediated by the GA receptor GA-INSENSITIVE DWARF1 (GID1) and DELLA proteins promotes hypocotyl elongation and inhibits anthocyanin accumulation. Whether CRY1 control of photomorphogenesis involves regulation of GA signaling is largely unknown. Here, we show that CRY1 signaling involves the inhibition of GA signaling through repression of GA-induced degradation of DELLA proteins. CRY1 physically interacts with DELLA proteins in a blue light-dependent manner, leading to their dissociation from SLEEPY1 (SLY1) and the inhibition of their ubiquitination. Moreover, CRY1 interacts directly with GID1 in a blue light-dependent but GA-independent manner, leading to the inhibition of the interaction between GID1 with DELLA proteins. These findings suggest that CRY1 controls photomorphogenesis through inhibition of GA-induced degradation of DELLA proteins and GA signaling, which is mediated by CRY1 inhibition of the interactions of DELLA proteins with GID1 and SCF^SLY1, respectively.

Blue light-dependent interactions of CRY1 with GID1 and DELLA proteins inhibit gibberellin (GA)-induced degradation of DELLA proteins to regulate GA signaling and photomorphogenesis.     

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.     

The rhythmic expression of genes controlling flowering time in southern and northern populations of invasive Ambrosia artemisiifolia

Aims Flowering time has been suggested to be an important adaptive trait during the dispersal of invasive species, and identifying the molecular mechanisms underlying flowering time may provide insight into the local adaptation during the process of invasion. Here, we conducted a preliminary exploration on the genetic basis of the differentiation of flowering time in Ambrosia artemisiifolia .Methods Using relative real-time fluorescent quantitative polymerase chain reaction, we investigated the expression levels of eight flowering-related genes, including AP1, FT, SOC1, CRY2, FKF1, GI, CO2 and SPY, in leaves and flowers at different time points in individuals from northern Beijing and southern Wuhan populations that exhibit significant differences in flowering times to identify any rhythmic changes in gene expression and their association with differential flowering times.Important findings The differentiation of flowering time in the A. artemisiifolia populations was closely associated with five genes involved in flowering pathways. The floral pathway integrators FT and SOC1 and floral meristem identity gene AP1 exhibited increased expression during flowering. The photoreceptor CRY2 in the light-dependent pathway and the SPY gene in the gibberellin pathway displayed specific expression patterns over time. In earlier-flowering Beijing plants, CRY2 expression was lower and SPY expression was higher than in Wuhan plants. The expression patterns of these five genes suggest a molecular basis for the differentiation of flowering time in A. artemisiifolia .     

Tomato contains homologues of Arabidopsis cryptochromes 1 and 2

Cryptochromes are blue light photoreceptors found in both plants and animals. They probably evolved from photolyases, which are blue/UV-light-absorbing photoreceptors involved in DNA repair. In seed plants, two different cryptochrome (CRY) genes have been found in Arabidopsis and one in Sinapis, while three genes have been found in the fern Adiantum. We report the characterisation of tomato CRY genes CRY1 and CRY2. They map to chromosomes 4 and 9, respectively, show relatively constitutive expression and encode proteins of 679 and 635 amino acids, respectively. These proteins show higher similarity to their Arabidopsis counterparts than to each other, suggesting that duplication between CRY1 and CRY2 is an ancient event in the evolution of seed plants. The seed plant cryptochromes form a group distinct from the fern cryptochromes, implying that only one gene was present in the common ancestor between these two groups of plants. Most intron positions in CRY genes from plants and ferns are highly conserved. Tomato cry1 and cry2 proteins carry C-terminal domains 210 and 160 amino acids long, respectively. Several conserved motifs are found in these domains, some of which are common to both types of cryptochromes, while others are cryptochrome-type-specific.