RESUMO
Spontaneous leaf color variation in bamboo provides the opportunity to study the mechanisms of leaf color formation and the breeding of ornamental bamboos. Despite the fact that many genes are known to be involved in leaf color variation in model plants, molecular mechanisms governing natural leaf color variation in bamboo have remained obscure. This study aimed to identify the genes responsible for the occurrence of such phenomena in bamboo using the suppression subtractive hybridization (SSH) method between green and albino leaves in Pseudosasa japonica f. A total of 1062 and 1004 differentially expressed transcripts were obtained from the forward and reverse SSH libraries, respectively. Subsequently, 59 differentially expressed unigenes with potential roles in leaf color formation, predicted via computational analysis of their functional relevance, were selected for further analysis using qPCR. Ten genes, involved in photosynthesis, plastid development, and cation signal transduction, showed 2-fold changes in expression levels between green and albino leaves. Further expression pattern analyses of these genes at three developmental stages revealed much lower expression abundance of Lhca1-encoded chlorophyll a/b binding protein in the albino leaves than in the green leaves. Our results suggest that, together with the concatenated negative pressure for subsequent photosynthetic processes, the albino phenotype is at least partly attributable to chloroplast inner membrane damage or to the impairment of photosynthetic pigment accumulation, which results from low Lhca1 expression.
Assuntos
Bambusa/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Pigmentação/genética , Folhas de Planta/genética , Bambusa/anatomia & histologia , Bambusa/crescimento & desenvolvimento , Clorofila/genética , Clorofila/metabolismo , Clorofila A , Proteínas de Ligação à Clorofila/genética , Proteínas de Ligação à Clorofila/metabolismo , Cloroplastos/genética , Cor , Fotossíntese , Folhas de Planta/anatomia & histologia , Folhas de Planta/crescimento & desenvolvimento , Transdução de Sinais , Técnicas de Hibridização SubtrativaRESUMO
TERMINAL FLOWER1 (TFL1) homologous genes play major roles in maintaining vegetative growth and inflorescence meristem characteristics in various plant species; however, to date, the function of the bamboo TFL1 homologous gene has not been described. In this study, a TFL1 homologous gene was isolated from Bambusa oldhamii and designated as BoTFL1-like. Phylogenetic analysis of TFL1 homologous genes revealed that BoTFL1-like shared more than 90% identity with the TFL1 genes of other Gramineae. RT-PCR analysis showed that the expression level of BoTFL1-like in floral buds was almost 3.5 times higher than in vegetative buds. In 35S::BoTFL1-like transgenic Arabidopsis thaliana plants, the time of flowering was significantly delayed by 5 to 9 days, and development of floral buds and sepals was severely affected compared to wild type Arabidopsis plants. This suggests that the BoTFL1-like gene may play roles in flowering time and flower morphological structure in B. oldhamii. The BoTFL1-like gene driven by the 35S promoter almost fully rescued the phenotype of the tfl1 mutant apart from the number of rosette inflorescences, indicating that the function of BoTFL1-like was similar to TFL1 in Arabidopsis. We conclude the TFL1 gene function has been conserved between B. oldhamii and A. thaliana.
Assuntos
Arabidopsis/genética , Bambusa/genética , Expressão Ectópica do Gene , Flores/genética , Mutação , Fenótipo , Proteínas de Plantas/genética , Sequência de Aminoácidos , Bambusa/classificação , Sequência de Bases , Clonagem Molecular , Modelos Moleculares , Dados de Sequência Molecular , Filogenia , Proteínas de Plantas/química , Conformação Proteica , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
The Neotropical woody bamboo genus Otatea is one of five genera in the subtribe Guaduinae. Of the eight described Otatea species, seven are endemic to Mexico and one is also distributed in Central and South America. Otatea acuminata has the widest geographical distribution of the eight species, and two of its recently collected populations do not match the known species morphologically. Parametric and non-parametric methods were used to delimit the species in Otatea using five chloroplast markers, one nuclear marker, and morphological characters. The parametric coalescent method and the non-parametric analysis supported the recognition of two distinct evolutionary lineages. Molecular clock estimates were used to estimate divergence times in Otatea. The results for divergence time in Otatea estimated the origin of the speciation events from the Late Miocene to Late Pleistocene. The species delimitation analyses (parametric and non-parametric) identified that the two populations of O. acuminata from Chiapas and Hidalgo are from two separate evolutionary lineages and these new species have morphological characters that separate them from O. acuminata s.s. The geological activity of the Trans-Mexican Volcanic Belt and the Isthmus of Tehuantepec may have isolated populations and limited the gene flow between Otatea species, driving speciation. Based on the results found here, I describe Otatea rzedowskiorum and Otatea victoriae as two new species, morphologically different from O. acuminata.
Assuntos
Bambusa/classificação , Bambusa/anatomia & histologia , Bambusa/genética , Teorema de Bayes , DNA de Cloroplastos/genética , Ecossistema , Fluxo Gênico , Especiação Genética , Funções Verossimilhança , México , Filogenia , Análise de Sequência de DNA , Estatísticas não ParamétricasRESUMO
Background: 4-coumarate:CoA ligase (4CL) plays an important role at the divergence point from general phenylpropanoid metabolism to several branch pathways. Although 4CL sin higher plants have been extensively studied, little has known about the 4CL gene of bamboo. Results: In current study, a Na4CL gene putative encoding 4-coumarate:CoA ligase (4CL) and its 5-flanking region were isolated from bamboo (Neosinocalamus affinis) by RACE-PCR and genomic DNA walker, respectively. Na4CL encodes a predicted protein of 557 amino acids, with conserved motifs of adenylate-forming enzymes. Phylogenetic analysis showed that Na4CL shared 62~85 percent identity with other known plant 4CLs, and cluster closely with some known 4CLs in monocots. Sequence analysis revealed conserved cis-acting elements (Box A and AC-II element) present in the Na4CL promoter. Additionally, a Na4CL RNAi construct was transformed into tobacco. Transgenic tobaccos displayed significant down-expression of endogenesis 4CL and reduced lignin contents. Conclusion: These results contribute to the knowledge of the presence of Na4CL gene and its possible role in phenylpropanoid metabolism.
Assuntos
Bambusa/genética , Clonagem Molecular , Ácidos Cumáricos , Coenzima A Ligases/genética , Bambusa/enzimologia , Interferência de RNA , Reação em Cadeia da Polimerase/métodos , Análise de SequênciaRESUMO
Fermented bamboo shoots are popular traditional food items of various ethnic groups of the northeastern India, especially in Manipur State. Dendrocalamus hamiltonii is an economically important bamboo species used to produce fermented bamboo shoots. We studied genetic variability of this bamboo species in Chandel and Imphal-East (commercial production districts), using AFLP molecular markers. Each of the selected primers detected polymorphisms and 1614 (95.8%) were found to be polymorphic. Cluster analysis based on Dice similarity coefficients using UPGMA differentiated the populations into two major groups. Principal coordinate analysis based on the AFLP data clearly separated the populations according to their genetic diversity and antioxidant activity. Four primers were tested through multiple regression analysis to identify marker-trait association between AFLP data and biochemical attributes, i.e., antioxidant activity and total cyanide content. The 273 bp generated by EcoRI-AAG(Joe)/MseI-CTC showed high positive correlation with antioxidant activity (r = 0.729, P < 0.01). The 396 bp generated by EcoRI-AAC(Ned)/MseI-CTG were negatively correlated with cyanide content (r = -0.694, P < 0.01). Thus, we found association of DNA markers with antioxidant activities and total cyanide content. These results could be of use for the identification of superior genotypes with desirable traits.
Assuntos
Análise do Polimorfismo de Comprimento de Fragmentos Amplificados/métodos , Bambusa/genética , Ecótipo , Variação Genética , Característica Quantitativa Herdável , Antioxidantes/farmacologia , Bambusa/anatomia & histologia , Bambusa/efeitos dos fármacos , Compostos de Bifenilo/metabolismo , Cianetos/metabolismo , Sequestradores de Radicais Livres/farmacologia , Marcadores Genéticos/genética , Geografia , Índia , Filogenia , Picratos/metabolismo , Extratos Vegetais/farmacologia , Polimorfismo Genético , Análise de Componente Principal , Análise de RegressãoRESUMO
PIF-like elements are the first-described members of a recently discovered and widespread superfamily of DNA transposons, named PIF/Harbinger. Complete and partial PIF-like elements have been isolated from hundreds of plant species. Previously, we identified 139 partial PIF-like transposases in the Bambusoideae, of which three were from the bamboo species Phyllostachys pubescens. Here we report identification and isolation of the first full-length PIF-like element (PpPIF-1) from P. pubescens; identification was made by chromosome walking, based on a modified magnetic enrichment procedure that allows efficient cloning of flanking sequences up to 3 kb in length. PpPIF-1 is 5953 bp in length, with 20-bp imperfect inverted terminal repeats and 3-bp target site duplications. This element contains two open reading frames, one encoding a putative transposase, including the complete DDE-domain typical of PIF/Harbinger elements from plants, and the other encoding a DNA-binding protein. There are seven termination codons and two frameshift mutations in the open reading frames, probably due to vertical inactivation.
Assuntos
Bambusa/genética , Genes de Plantas , Sequência de Aminoácidos , Sequência de Bases , Passeio de Cromossomo , Primers do DNA , Mutação da Fase de Leitura , Dados de Sequência Molecular , Fases de Leitura Aberta , Filogenia , Homologia de Sequência de Aminoácidos , Transposases/química , Transposases/genéticaRESUMO
Phyllostachys pubescens is a woody bamboo with the highest ecological, economic, and cultural values of all bamboos in Asia. There is more genomic data available for P. pubescens than for any other bamboo species, including 2.12-Mb genome survey sequences (GSS) and 11.4-Mb full-length cDNA sequences (FL-cDNAs) currently deposited in GenBank. Analysis of these sequences revealed that transposable elements (TEs) are abundant, diverse and polyphyletic in the P. pubescens genome, of which Ty3-gypsy and Ty1-copia are the two most abundant families. Phylogenic analysis showed that both elements probably arose before the Bambusoideae separated from the other Poaceae subfamilies. We found evidence that the distribution of some intragenic TEs correlated with transcript profiles, of which Mutator elements preferred to insert in the transcripts of transcription factors. Additionally, we found that the abundance of SSRs in TEs (4.56%) was significantly higher than in GSS (0.098%) and in FL-cDNAs (2.60%) in P. pubescens genome, and TA/AT and CT/AG repeats were found to be intimately associated with En/Spm and Mutator elements, respectively. Our data provide a glimpse of the structure and evolution of P. pubescens genome, although large-scale sequencing of the genome would be required to fully understand the architecture of the P. pubescens genome.
Assuntos
Bambusa/genética , DNA Complementar/genética , Genoma de Planta , Genômica/métodos , Repetições de Microssatélites , Biologia de Sistemas/métodos , Elementos de DNA Transponíveis , Bases de Dados Genéticas , Evolução Molecular , Filogenia , Análise de Sequência de DNA , Fatores de Transcrição/genéticaRESUMO
RNA isolation from hard and woody internodal bamboo (Bambusa balcooa) tissue is very difficult due to the presence of secondary metabolites, polysaccharides, and polyphenolics. These compounds often co-precipitate with isolated RNA and hinder downstream applications. We have developed an efficient, cost effective and reproducible RNA isolation method from hard tissue of bamboo internode. This protocol includes an additional organic solvent refinement steps to remove endogenous phenolic compounds and acidic phenol (pH 4.2) to critically stabilize RNA in extraction buffer. In addition to these, two 2M Lithium chloride washing steps were introduced to eliminate DNA and polysaccharides contamination. The RNA isolated from the present protocol was found to be superior, when compared to total RNA extracted by other available protocols. The A260/A280 absorption ratio of the isolated RNA was found ranging between 1.89-1.97. The integrity of 28S and 18S rRNA was highly satisfactory when analyzed in agarose denaturing gel. RNA was further used for RT PCR, northern hybridization, cDNA library and subtractive hybridization without any further refinement.
Assuntos
RNA de Plantas/isolamento & purificação , Bambusa/genética , Northern Blotting , Compostos Fenólicos , Reação em Cadeia da Polimerase , PolissacarídeosRESUMO
Unlike other plants, bamboo (Bambusoideae) flowering is an elusive physiological phenomena, because it is unpredictable, long-periodic, gregarious, and uncontrollable; also, bamboo plants usually die after flowering. The flowering mechanism in Arabidopsis thaliana, a eudicot model species, is well established, but it remains unknown in bamboo species. We found 4470 and 3878 expressed sequence tags in the flower bud and vegetative shoot cDNA libraries, respectively, of the bamboo species, Bambusa oldhamii. Different genes were found expressed in bamboo flower buds compared to vegetative shoots, based on the Munich Information Center for Protein Sequences functional categorization; flowering-related genes were also identified in this species. We also identified Arabidopsis flowering-specific homologs that are involved in its photoperiod in this bamboo species, along with autonomous, vernalization and gibberellin-dependent pathways, indicating that bamboos may have a similar mechanism to control floral transition. Some bamboo expressed sequence tags shared high similarity with those of rice, but others did not match any known sequences. Our data lead us to conclude that bamboo may have its own unique flowering genes. This information can help us understand bamboo flowering and provides useful experimental methods to study the mechanisms involved.