Project description:'Systems-wide' approaches such as microarray RNA-profiling are ideally suited to the study of the complex, overlapping responses of plants to biotic and abiotic stresses. However, commercial microarrays are only available for a limited number of plant species and development costs are so substantial as to be prohibitive for most research groups. Here, we evaluate the use of cross-hybridisation to Affymetrix oligonucleotide GeneChip microarrays to profile the response of the banana (Musa sp.) leaf transcriptome to drought stress using a genomic DNA (gDNA)-based probe-selection strategy to improve the efficiency of detection of differentially expressed Musa transcripts. GeneChip analyses were performed to profile the response of the banana (Musa sp.) leaf transcriptome to drought stress using a genomic DNA (gDNA)-based probe-selection strategy to improve the efficiency of detection of differentially expressed Musa transcripts.
Project description:'Systems-wide' approaches such as microarray RNA-profiling are ideally suited to the study of the complex, overlapping responses of plants to biotic and abiotic stresses. However, commercial microarrays are only available for a limited number of plant species and development costs are so substantial as to be prohibitive for most research groups. Here, we evaluate the use of cross-hybridisation to Affymetrix oligonucleotide GeneChip microarrays to profile the response of the banana (Musa sp.) leaf transcriptome to drought stress using a genomic DNA (gDNA)-based probe-selection strategy to improve the efficiency of detection of differentially expressed Musa transcripts.
Project description:Small RNAs (21-24 nt) are pivotal regulators of gene expression that guide both transcriptional and post-transcriptional silencing mechanisms in diverse eukaryotes, including most if not all plants. MicroRNAs (miRNAs) and short interfering RNAs (siRNAs) are the two major types, both of which have a demonstrated and important role in plant development, stress responses and pathogen resistance. In this work, we used a deep sequencing approach (Sequencing-By-Synthesis, or SBS) to develop sequence resources of small RNAs from Musa acuminata tissues (including leaves, male flowers and immature fruit). The high depth of the resulting datasets enabled us to examine in detail critical small RNA features as size distribution, tissue-specific regulation and sequence conservation between different organs in this species. We also developed database resources and a dedicated website (http://smallrna.udel.edu/) with computational tools for allowing other users to identify new miRNAs or siRNAs involved in specific regulatory pathways, verify the degree of conservation of these sequences in other plant species and map small RNAs on genes or larger regions of the maize genome under study.
Project description:Small RNAs (21-24 nt) are pivotal regulators of gene expression that guide both transcriptional and post-transcriptional silencing mechanisms in diverse eukaryotes, including most if not all plants. MicroRNAs (miRNAs) and short interfering RNAs (siRNAs) are the two major types, both of which have a demonstrated and important role in plant development, stress responses and pathogen resistance. In this work, we used a deep sequencing approach (Sequencing-By-Synthesis, or SBS) to develop sequence resources of small RNAs from Musa acuminata tissues (including leaves, male flowers and immature fruit). The high depth of the resulting datasets enabled us to examine in detail critical small RNA features as size distribution, tissue-specific regulation and sequence conservation between different organs in this species. We also developed database resources and a dedicated website (http://smallrna.udel.edu/) with computational tools for allowing other users to identify new miRNAs or siRNAs involved in specific regulatory pathways, verify the degree of conservation of these sequences in other plant species and map small RNAs on genes or larger regions of the maize genome under study. Small RNA libraries were derived from leaves, male flowers and immature fruit of Musa acuminata. Total RNA was isolated using the Plant RNA Purification Reagent (Invitrogen), and submitted to Illumina (Hayward, CA, http://www.illumina.com) for small RNA library construction using approaches described in (Lu et al., 2007) with minor modifications. The small RNA libraries were sequenced with the Sequencing-By-Synthesis (SBS) technology by Illumina. PERL scripts were designed to remove the adapter sequences and determine the abundance of each distinct small RNA. We thank Nicolas Roux for providing the plant material as well as Kan Nobuta and Gayathri Mahalingam for assistance with the computational methods.
Project description:Bananas and plantains are highly susceptible to drought and may lead to yield reductions of up to 65%. Musa balbisiana harbors many biotic and abiotic stress-tolerant characteristics and there is urgent need to integrate B-genomic regions into cultivars for various stress resistance, nutrition, and vigor characteristics in the new banana varieties. With the above objective GBS technology was applied to obtain SNPs on a unique population developed which was obtained crossing Musa acuminata and Musa balbisiana. Here, in the current submission we deposit the SNPs identified in the mentioned population (AAxBB).
Project description:Cold-sensitive Cavendish Banana and relatively cold-tolerant Dajiao (Musa spp.) comprise an important part of diets for millions of people around the globe. Low temperature is one of the key environment stresses which greatly affect the global banana production. However, little is known about the changes of global protein phosphorylation in Musa spp. and their regulatory roles in response to cold stress. In this study, we employed a TMT6-plex quantitative analysis to conduct a global phosphoproteome profiling between Cavendish Banana and Dajiao subject to the cold stress for 0 hour and 3 hour. A total of 679 phosphopeptides containing 772 distinct phosphorylated sites from 529 phosphoproteins were identified in Cavendish Banana, 180 phosphorylation sites (belonging to 147 phosphoproteins) were differentially changed after 3 h cold stress. While in Dajiao 241 phosphopeptides with 271 individual phosphosites from 207 phosphoproteins were confidently identified, and 83 phosphorylation sites from 63 phosphoproteins were differentially changed under 3 h cold stress. Bioinformatic analysis of protein interaction network indicated that Mitogen-activated protein kinase kinase 2 (MKK2) was located in the center of the MAPK signaling network along with 7 other members whose phosphorylated site abundances were remarkably differentiated between Cavendish Banana and Dajiao in response to cold stress. Western blotting of MKK2 protein and its T31 phosphorylated site showed the increased expression of MKK2 in the time course of cold stress, with no detectable T31 phosphorylation in Cavendish Banana. On the contrary, the decreased MKK2 expression with increased T31 phosphorylation was consistently observed in Dajiao. These results suggest that the MKK2 interaction network in Dajiao, along with other cold-specific phosphoproteins found in this study, appears to play an important role in the molecular mechanisms of Dajiao being high tolerance to cold stress. The results also provide new evidence that cellular MKK2 phosphorylation as a signaling pathway plays an important role in abiotic stress tolerance that serves as a universal plant cold tolerance mechanism. To the best of our knowledge, this is the first report of MKK2 network involved in the regulatory of the Musa spp. response to cold stress.