RESUMO
Eruca vesicaria subsp sativa is one of the most tolerant Cruciferae species to drought, and dehydration-responsive element-binding protein 2A (DREB2A) is involved in responses to salinity, heat, and particularly drought. In this study, a gene encoding EvDREB2A was cloned and characterized in E. vesicaria subsp sativa. The full-length EvDREB2A cDNA sequence contained a 388-bp 5'-untranslated region (UTR), a 348-bp 3'-UTR, and a 1002-bp open reading frame that encoded 334 amino acid residues. The theoretical isoelectric point of the EvDREB2A protein was 4.80 and the molecular weight was 37.64 kDa. The genomic sequence of EvDREB2A contained no introns. Analysis using SMART indicated that EvDREB2A contains a conserved AP2 domain, similar to other plant DREBs. Phylogenetic analysis revealed that EvDREB2A and DREB2As from Brassica rapa, Eutrema salsugineum, Arabidopsis thaliana, Arabidopsis lyrata, and Arachis hypogaea formed a small subgroup, which clustered with DREB2Bs from A. lyrata, A. thaliana, Camelina sativa, and B. rapa to form a larger subgroup. EvDREB2A is most closely related to B. rapa DREB2A, followed by DREB2As from E. salsugineum, A. thaliana, A. hypogaea, and A. lyrata. A quantitative real-time polymerase chain reaction indicated that EvDREB2A expression was highest in the leaves, followed by the roots and hypocotyls, and was lowest in the flower buds. EvDREB2A could be used to improve drought tolerance in crops.
Assuntos
Brassicaceae/genética , Proteínas de Plantas/genética , Clonagem Molecular , DNA Complementar/química , DNA Complementar/genética , Regulação da Expressão Gênica de Plantas , Folhas de Planta/genética , Proteínas de Plantas/química , Raízes de Plantas/genética , Domínios e Motivos de Interação entre Proteínas , Análise de Sequência de DNARESUMO
Nicotianamine (NA) is a ubiquitous metabolite in plants that bind heavy metals, is crucial for metal homeostasis, and is also an important metal chelator that facilitates long-distance metal transport and sequestration. NA synthesis is catalyzed by the enzyme nicotianamine synthase (NAS). Eruca vesicaria subsp sativa is highly tolerant to Ni, Pb, and Zn. In this study, a gene encoding EvNAS was cloned and characterized in E. vesicaria subsp sativa. The full-length EvNAS cDNA sequence contained a 111-bp 5'-untranslated region (UTR), a 155-bp 3'-UTR, and a 966-bp open reading frame encoding 322-amino acid residues. The EvNAS genomic sequence contained no introns, which is similar to previously reported NAS genes. The deduced translation of EvNAS contained a well-conserved NAS domain (1-279 amino acids) and an LIKI-CGEAEG box identical to some Brassica NAS and to the LIRL-box in most plant NAS, which is essential for DNA binding. Phylogenetic analysis indicated that EvNAS was most closely related to Brassica rapa NAS3 within the Cruciferae, followed by Thlaspi NAS1, Camelina NAS3, and Arabidopsis NAS3. A reverse transcription-polymerase chain reaction indicated that EvNAS expression was greatest in the leaves, followed by the flower buds and hypocotyls. EvNAS was moderately expressed in the roots.
Assuntos
Alquil e Aril Transferases/genética , Sequência de Aminoácidos/genética , Brassicaceae/enzimologia , Filogenia , Alquil e Aril Transferases/biossíntese , Clonagem Molecular , DNA Complementar/genética , Regulação da Expressão Gênica de Plantas , Ferro/metabolismo , Folhas de Planta/genética , Raízes de PlantasRESUMO
The formation of polyploid was mainly through the combination of 2n gamete, and the apomixis played an important role in plant evolution and plant breeding. In general, the formation of male gamete and female gamete in apomictic plants are obviously different, i.e., the formation of embryo sac was through apomixis (form into 2n female gamete); however, the formation of male gamete was through normal meiosis (form into 1n male gamete). The 2n male gamete apomictic wheat grass was more important than 1n male gamete apomictic wheat grass, because the 2n male gamete can bring all genes including apomictic gene into other plant. The meiosis of 2n male gamete apomictic wheat grass (Elymus. rectisetus) was observed through Olympus AH3 microscope; the results showed that the meiosis of 2n gamete apomictic wheat grass was very abnomal. In the interphase, lots of mini-nucleus were found. Inverted cycle, multibivalent and lagging chromosome were also found in the prophase. In metaphase, chromosomes frequently show unequal division tendency; In anaphase I and telophase I, completely unequal division, modified meiosis, the tendency of nucleus fusion and other abnormal phenomena were found. These phenomena directly lead to the production of 2n gamete. In meiosis II, the abnormalities mainly lie in the proceeding of microspore, dyad (55.7%), triad (23.7%) and tetrad (21.6%) were found. The abnormal of those proceeding also lead to the production of 2n gamete.