Cultivar-specific markers, mutations, and chimerisim of Cavendish banana somaclonal variants resistant to Fusarium oxysporum f. sp. cubense tropical race 4.

BACKGROUND: The selection of tissue culture-derived somaclonal variants of Giant Cavendish banana (Musa spp., Cavendish sub-group AAA) by the Taiwan Banana Research Institute (TBRI) has resulted in several cultivars resistant to Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4), a destructive fungus threatening global banana production. However, the mutations in these somaclonal variants have not yet been determined. We performed an RNA-sequencing (RNA-seq) analysis of three TBRI Foc TR4-resistant cultivars: 'Tai-Chiao No. 5' (TC5), 'Tai-Chiao No. 7' (TC7), and 'Formosana' (FM), as well as their susceptible progenitor 'Pei-Chiao' (PC), to investigate the sequence variations among them and develop cultivar-specific markers. RESULTS: A group of single-nucleotide variants (SNVs) specific to one cultivar were identified from the analysis of RNA-seq data and validated using Sanger sequencing from genomic DNA. Several SNVs were further converted into cleaved amplified polymorphic sequence (CAPS) markers or derived CAPS markers that could identify the three Foc TR4-resistant cultivars among 6 local and 5 international Cavendish cultivars. Compared with PC, the three resistant cultivars showed a loss or alteration of heterozygosity in some chromosomal regions, which appears to be a consequence of single-copy chromosomal deletions. Notably, TC7 and FM shared a common deletion region on chromosome 5; however, different TC7 tissues displayed varying degrees of allele ratios in this region, suggesting the presence of chimerism in TC7. CONCLUSIONS: This work demonstrates that reliable SNV markers of tissue culture-derived and propagated banana cultivars with a triploid genome can be developed through RNA-seq data analysis. Moreover, the analysis of sequence heterozygosity can uncover chromosomal deletions and chimerism in banana somaclonal variants. The markers obtained from this study will assist with the identification of TBRI Cavendish somaclonal variants for the quality control of tissue culture propagation, and the protection of breeders' rights.

Overexpression of a Novel ERF-X-Type Transcription Factor, OsERF106MZ, Reduces Shoot Growth and Tolerance to Salinity Stress in Rice.

Transcription factors (TFs) such as ethylene-responsive factors (ERFs) are important for regulating plant growth, development, and responses to abiotic stress. Notably, more than half of the rice ERF-X group members, including ethylene-responsive factor 106 (OsERF106), are abiotic stress-responsive genes. However, their regulatory roles in abiotic stress responses remain poorly understood. OsERF106, a salinity-induced gene of unknown function, is annotated differently in RAP-DB and MSU RGAP. In this study, we isolated a novel (i.e., previously unannotated) OsERF106 gene, designated OsERF106MZ (GenBank accession No. MZ561461), and investigated its role in regulating growth and the response to salinity stress in rice. OsERF106MZ is expressed in germinating seeds, primary roots, and developing flowers. Overexpression of OsERF106MZ led to retardation of growth, relatively high levels of both malondialdehyde (MDA) and reactive oxygen species (ROS), reduced catalase (CAT) activity, and overaccumulation of both sodium (Na+) and potassium (K+) ions in transgenic rice shoots. Additionally, the expression of OsHKT1.3 was downregulated in the shoots of transgenic seedlings grown under both normal and NaCl-treated conditions, while the expression of OsAKT1 was upregulated in the same tissues grown under NaCl-treated conditions. Further microarray and qPCR analyses indicated that the expression of several abiotic stress-responsive genes such as OsABI5 and OsSRO1c was also altered in the shoots of transgenic rice grown under either normal or NaCl-treated conditions. The novel transcription factor OsERF106MZ negatively regulates shoot growth and salinity tolerance in rice through the disruption of ion homeostasis and modulation of stress-responsive gene expression.

The nucleolar protein SAHY1 is involved in pre-rRNA processing and normal plant growth.

Although the nucleolus is involved in ribosome biogenesis, the functions of numerous nucleolus-localized proteins remain unclear. In this study, we genetically isolated Arabidopsis thaliana salt hypersensitive mutant 1 (sahy1), which exhibits slow growth, short roots, pointed leaves, and sterility. SAHY1 encodes an uncharacterized protein that is predominantly expressed in root tips, early developing seeds, and mature pollen grains and is mainly restricted to the nucleolus. Dysfunction of SAHY1 primarily causes the accumulation of 32S, 18S-A3, and 27SB pre-rRNA intermediates. Coimmunoprecipitation experiments further revealed the interaction of SAHY1 with ribosome proteins and ribosome biogenesis factors. Moreover, sahy1 mutants are less sensitive to protein translation inhibitors and show altered expression of structural constituents of ribosomal genes and ribosome subunit profiles, reflecting the involvement of SAHY1 in ribosome composition and ribosome biogenesis. Analyses of ploidy, S-phase cell cycle progression, and auxin transport and signaling indicated the impairment of mitotic activity, translation of auxin transport carrier proteins, and expression of the auxin-responsive marker DR5::GFP in the root tips or embryos of sahy1 plants. Collectively, these data demonstrate that SAHY1, a nucleolar protein involved in ribosome biogenesis, plays critical roles in normal plant growth in association with auxin transport and signaling.

Genetic analyses of the interaction between abscisic acid and gibberellins in the control of leaf development in Arabidopsis thaliana.

Although abscisic acid (ABA) and gibberellins (GAs) play pivotal roles in many physiological processes in plants, their interaction in the control of leaf growth remains elusive. In this study, genetic analyses of ABA and GA interplay in leaf growth were performed in Arabidopsis thaliana. The results indicate that for the ABA and GA interaction, leaf growth of both the aba2/ga20ox1 and aba2/GA20ox1 plants, which were derived from the crosses of aba2×ga20ox1 and aba2×GA20ox1 overexpressor, respectively, exhibits partially additive effects but is similar to the aba2 mutant. Consistently, the transcriptome analysis suggests that a substantial proportion (45-65%) of the gene expression profile of aba2/ga20ox1 and aba2/GA20ox1 plants overlap and share a pattern similar to the aba2 mutant. Thus, these data suggest that ABA deficiency dominates leaf growth regardless of GA levels. Moreover, the gene ontology (GO) analysis indicates gene enrichment in the categories of hormone response, developmental and metabolic processes, and cell wall organization in these three genotypes. Leaf developmental genes are also involved in the ABA-GA interaction. Collectively, these data support that the genetic relationship of ABA and GA interaction involves multiple coordinated pathways rather than a simple linear pathway for the regulation of leaf growth.

The Arabidopsis short-chain dehydrogenase/reductase 3, an abscisic acid deficient 2 homolog, is involved in plant defense responses but not in ABA biosynthesis.

ABSCISIC ACID DEFICIENT2 (ABA2) encodes a short-chain dehydrogenase/reductase1 (SDR1) that catalyzes the multi-step conversion of xanthoxin to abscisic aldehyde during abscisic acid (ABA) biosynthesis in Arabidopsis thaliana. In this study, AtSDR2 and AtSDR3, the two closest homologs to AtABA2, were investigated for their potential role in ABA biosynthesis. AtSDR2 showed undetectable transcription in plants grown under normal conditions or under stress. AtSDR3 and AtABA2 have different spatial and temporal expression patterns. Complementation testing demonstrated that the pABA2::SDR3 transgene failed to complement the aba2 mutant phenotype, and that transgenic plants showed the same levels of ABA as the aba2 mutants. These data suggest that AtSDR3 confers no functional redundancy to AtABA2 in ABA biosynthesis. Interestingly, microarray data derived from Genevestigator suggested that AtSDR3 might have a function that is related to plant defense. Pseudomonas syringae pv. tomato (Pst) DC3000 infection and systemic acquired resistance (SAR) activator application further demonstrated that AtSDR3 plays an important role in plant defense responses at least partially through the regulation of AtPR-1 gene expression.

Antagonism between abscisic acid and ethylene in Arabidopsis acts in parallel with the reciprocal regulation of their metabolism and signaling pathways.

Although abscisic acid (ABA) and ethylene have antagonistic functions in the control of plant growth and development, including seed germination and early seedling development, it remains unknown whether a convergent point exists between these two signaling pathways or whether they operate in parallel in Arabidopsis thaliana. To elucidate this issue, four ethylene mutants, ctr1, ein2, ein3, and ein6, were crossed with aba2 (also known as gin1-3) to generate double mutants. Genetic epistasis analysis revealed that all of the resulting double mutants displayed aba2 mutant phenotypes with a small plant size and wiltiness when grown in soil or on agar plates. Further ethylene sensitivity or triple response analyses demonstrated that these double mutants also retained the ctr1 or ein mutant phenotypes, showing ethylene constitutive triple and insensitive responses, respectively. Our current data therefore demonstrate that ABA and ethylene act in parallel, at least in primary signal transduction pathways. Moreover, by microarray analysis we found that an ACC oxidase (ACO) was significantly upregulated in the aba2 mutant, whereas the 9-CIS-EPOXYCAROTENOID DIOXYGENASE 3 (NCED3) gene in ein2 was upregulated, and both the ABSCISIC ACID INSENSITIVE1 (ABI1) and cytochrome P450, family 707, subfamily A, polypeptide 2 (CYP707A2) genes in etr1-1 were downregulated. These data further suggest that ABA and ethylene may control the hormonal biosynthesis, catabolism, or signaling of each other to enhance their antagonistic effects upon seed germination and early seedling growth.