Bamboo are woody grass species containing important economic and ecological values. Lei bamboo (Phyllostachys violascens) is a kind of shoot-producing bamboo species with the highest economic yield per unit area. However, identifying different varieties of Lei bamboo based on morphological characteristics is difficult. Microsatellites play an important role in plant identification and genetic diversity analysis and are superior to other molecular markers. In this study, we identified 18,356 expressed sequence tag-simple sequence repeat (EST-SSR) loci in Lei bamboo transcriptome data. A total of 11,264 primer pairs were successfully designed from unigenes of all EST-SSR loci, and 96 primer pairs were randomly selected and synthesized. A total of 54 primer pairs were used for classifying 16 Lei bamboo varieties and 10 different Phyllostachys species. The number of polymorphism alleles among the 54 primer pairs ranged from 3 to 12 for P. violascens varieties and 3 to 20 for Phyllostachys. The phylogenetic tree based on polymorphism alleles successfully distinguished 16 P. violascens varieties and 10 Phyllostachys species. Our study provides abundant EST-SSR resources that are useful for genetic diversity analysis and molecular verification of bamboo and suggests that SSR markers developed from Lei bamboo are more efficient and reliable than ISSR, SRAP or AFLP markers.
Bamboo is known for its edible shoots and beautiful texture and has considerable economic and ornamental value. Unique among traditional flowering plants, many bamboo plants undergo extensive synchronized flowering followed by large-scale death, seriously affecting the productivity and application of bamboo forests. To date, the molecular mechanism of bamboo flowering characteristics has remained unknown. In this study, a SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1)-like gene, BoMADS50, was identified from Bambusa oldhamii. BoMADS50 was highly expressed in mature leaves and the floral primordium formation period during B. oldhamii flowering and overexpression of BoMADS50 caused early flowering in transgenic rice. Moreover, BoMADS50 could interact with APETALA1/FRUITFULL (AP1/FUL)-like proteins (BoMADS14-1/2, BoMADS15-1/2) in vivo, and the expression of BoMADS50 was significantly promoted by BoMADS14-1, further indicating a synergistic effect between BoMADS50 and BoAP1/FUL-like proteins in regulating B. oldhamii flowering. We also identified four additional transcripts of BoMADS50 (BoMADS50-1/2/3/4) with different nucleotide variations. Although the protein-CDS were polymorphic, they had flowering activation functions similar to those of BoMADS50. Yeast one-hybrid and transient expression assays subsequently showed that both BoMADS50 and BoMADS50-1 bind to the promoter fragment of itself and the SHORT VEGETATIVE PHASE (SVP)-like gene BoSVP, but only BoMADS50-1 can positively induce their transcription. Therefore, nucleotide variations likely endow BoMADS50-1 with strong regulatory activity. Thus, BoMADS50 and BoMADS50-1/2/3/4 are probably important positive flowering regulators in B. oldhamii. Moreover, the functional conservatism and specificity of BoMADS50 and BoMADS50-1 might be related to the synchronized and sporadic flowering characteristics of B. oldhamii.
NAC (NAM, AFAT, and CUC) proteins play necessary roles in plant response to environmental stresses. However, the functional roles of NAC genes in moso bamboo (Phyllostachys edulis), an essential economic perennial woody bamboo species, are not well documented. In this study, we retrieved 152 PeNAC genes from the moso bamboo V2 genome, and PeSNAC-1 was isolated and functionally characterized. PeSNAC-1 was localized in the nucleus and had no transactivation activity in yeast. PeSNAC-1 extremely expressed in rhizome and young roots (0.1 and 0.5 cm) and was significantly induced by drought and salt treatments but repressed by abscisic acid (ABA), methyl jasmonate (MeJA) and high temperature (42°C) in moso bamboo. Under water shortage and salinity conditions, survival ratios, Fv/Fm values, physiological indexes such as activities of superoxide dismutase, peroxidase, and catalase and contents of malondialdehyde, H2O2, and proline were significantly higher in transgenic rice than the wild type, which suggests enhanced tolerance to drought and salt stress in PeSANC-1 overexpressed plants. Transcript levels of Na+/H+ antiporter and Na+ transporter genes (OsSOS1, OsNHX1 and OsHKT1;5), ABA signaling and biosynthesis genes (OsABI2, OsRAB16, OsPP2C68, OsLEA3–1, OsLEA3, OsNCED3, OsNCED4 and OsNCED5) and ABA independent genes (OsDREB1A, OsDREB1B and OsDREB2A) were substantially higher in transgenic as compared to the wild type. Moreover, protein interaction analysis revealed that PeSNAC-1 could interact with stress responsive PeSNAC-2/4 and PeNAP-1/4/5 in both yeast and plant cells, which indicates a synergistic effect of those proteins in regulating the moso bamboo stress response. Our data demonstrate that PeSNAC-1 likely improved salt and drought stress tolerance via modulating gene regulation in both ABA dependent and independent signaling pathways in transgenic rice. In addition, PeSNAC-1 functions as an important positive stress regulator in moso bamboo, participating in PeSNAC-1 and PeSNAC-2/−4 or PeSNAC-1 and PeNAP-1/4/5 interaction networks.
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