Phytohormone abscisic acid (ABA) regulates many aspects of plant development and growth. To explore the molecular mechanism of ABA, we identified the novel ABA-regulated genes in Arabidopsis thaliana by searching for genes possessing two or more ABREs (ABA-responsive elements). One of these genes, two or more ABREs-containing gene 2 (TMAC2) is highly induced by ABA and NaC1. Database searches revealed that TMAC2 encodes a protein with no domains of known function. Expression of TMAC2-GFP fusion protein in Arabidopsis mesophyll protoplasts indicated that TMAC2 is targeted to the nucleus. Although the gene has a basal level of expression in various Arabidopsis organs/tissues except for adult leaves, a high expression level was detected in roots. Constitutive overexpression of TMAC2 in plants resulted in the insensitivity to ABA and NaCl, suggesting that TMAC2 plays a negative role in ABA and salt stress responses. Furthermore, TMAC2-overexpressing plants exhibited the short roots, late flowering and starch-excess phenotypes. RT-PCR analysis showed that decreased expression of two floral- and one starch degradation-related genes, SOC1/AGL20 and SEP3/AGL9, and SEX1, respectively, may lead to altered phenotypes of TMAC2-overexpressing plants. Taken together, our data reveal that TMAC2 acts in the nucleus and is an important negative regulator of ABA and salt stress responses, and could play a critical role in controlling root elongation, floral initiation and starch degradation.

To investigate the organization of and mode of selection in the α-tubulin genes, full-length α-tubulin genes were cloned from four intraspecific taxa of Miscanthus sinensis and its close relatives M. floridulus and M. condensatus using standard polymerase chain reaction (PCR) and rapid amplification of genomic ends (RAGE)-PCR strategies. Genealogical analysis of angiosperms recovered a monophyletic group of Miscanthus α-tubulin genes, which is homologous to the tua5 locus of maize. Two clusters of nearly equal frequency revealed paraphyly within each Miscanthus taxon. Between-cluster recombination was frequent. Additional evidence for co-occurrence of two haplotypes within individuals and a large-scale crossover all suggested a likely allelic relationship between the Miscanthus clusters. Given a long between-species divergence time in Miscanthus, wide occurrence of the trans-species polymorphisms in α-tubulin genes and the approximately equal frequency of each allelic type make it extremely unlikely that α-tubulin diversity has been maintained under neutrality. Balancing selection may have contributed to such an apportioning of genetic variability as well as to high levels of genetic variation in α-tubulin and higher substitution rates at synonymous sites of exons than at intron bases of M. sinensis. In addition, certain effects of demographic oscillation may have distorted the scenario of a functional locus operating under balancing selection.

Phalaenopsis spp. represent the most popular orchids worldwide. Both P. equestris and P. aphrodite are the two important breeding parents with the whole genome sequence available. However, marker–trait association is rarely used for floral traits in Phalaenopsis breeding. Here, we analyzed markers associated with aesthetic traits of Phalaenopsis orchids by using genome-wide association study (GWAS) with the F1 population P. Intermedia of 117 progenies derived from the cross between P. aphrodite and P. equestris. A total of 113,517 single nucleotide polymorphisms (SNPs) were identified in P. Intermedia by using genotyping-by-sequencing with the combination of two different restriction enzyme pairs, Hinp1 I/Hae III and Apek I/Hae III. The size-related traits from flowers were negatively related to the color-related traits. The 1191 SNPs from Hinp1 I/ Hae III and 23 simple sequence repeats were used to establish a high-density genetic map of 19 homolog groups for P. equestris. In addition, 10 quantitative trait loci were highly associated with four color-related traits on chromosomes 2, 5 and 9. According to the sequence within the linkage disequilibrium regions, 35 candidate genes were identified and related to anthocyanin biosynthesis. In conclusion, we performed marker-assisted gene identification of aesthetic traits with GWAS in Phalaenopsis orchids.

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