Global warming is predicted to have a general negative effect on plant growth due to the damaging effect of high temperatures on plant development. The increasing threat of climatological extremes including very high temperatures might lead to catastrophic loss of crop productivity and result in wide spread famine. In this review, we assess the impact of global climate change on the agricultural crop production. There is a differential effect of climate change both in terms of geographic location and the crops that will likely show the most extreme reductions in yield as a result of expected extreme fluctuations in temperature and global warming in general. High temperature stress has a wide range of effects on plants in terms of physiology, biochemistry and gene regulation pathways. However, strategies exist to crop improvement for heat stress tolerance. In this review, we present recent advances of research on all these levels of investigation and focus on potential leads that may help to understand more fully the mechanisms that make plants tolerant or susceptible to heat stress. Finally, we review possible procedures and methods which could lead to the generation of new varieties with sustainable yield production, in a world likely to be challenged both by increasing population, higher average temperatures and larger temperature fluctuations.

Plant tolerance to high temperature in a changing environment: scientific fundamentals and production of heat stress-tolerant crops

Water limitation affects all types of organisms at some stage during their life cycle; therefore, many strategies have been selected through evolution to cope with water deficit, including changes in enzyme activities and in gene expression, among others. In plants, a group of very hydrophilic

The enigmatic LEA proteins and other hydrophilins.

The size and complexity of conifer genomes has, until now, prevented full genome sequencing and assembly. The large research community and economic importance of loblolly pine, Pinus taeda L., made it an early candidate for reference sequence determination. We develop a novel strategy to sequence the genome of loblolly pine that combines unique aspects of pine reproductive biology and genome assembly methodology. We use a whole genome shotgun approach relying primarily on next generation sequence generated from a single haploid seed megagametophyte from a loblolly pine tree, 20-1010, that has been used in industrial forest tree breeding. The resulting sequence and assembly was used to generate a draft genome spanning 23.2 Gbp and containing 20.1 Gbp with an N50 scaffold size of 66.9 kbp, making it a significant improvement over available conifer genomes. The long scaffold lengths allow the annotation of 50,172 gene models with intron lengths averaging over 2.7 kbp and sometimes exceeding 100 kbp in length. Analysis of orthologous gene sets identifies gene families that may be unique to conifers. We further characterize and expand the existing repeat library based on the de novo analysis of the repetitive content, estimated to encompass 82% of the genome. In addition to its value as a resource for researchers and breeders, the loblolly pine genome sequence and assembly reported here demonstrates a novel approach to sequencing the large and complex genomes of this important group of plants that can now be widely applied.

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Decoding the massive genome of loblolly pine using haploid DNA and novel assembly strategies

BackgroundThe size and complexity of conifer genomes has, until now, prevented full genome sequencing and assembly. The large research community and economic importance of loblolly pine, Pinus taeda L., made it an early candidate for reference sequence determination.ResultsWe develop a novel strategy to sequence the genome of loblolly pine that combines unique aspects of pine reproductive biology and genome assembly methodology. We use a whole genome shotgun approach relying primarily on next generation sequence generated from a single haploid seed megagametophyte from a loblolly pine tree, 20-1010, that has been used in industrial forest tree breeding. The resulting sequence and assembly was used to generate a draft genome spanning 23.2 Gbp and containing 20.1 Gbp with an N50 scaffold size of 66.9 kbp, making it a significant improvement over available conifer genomes. The long scaffold lengths allow the annotation of 50,172 gene models with intron lengths averaging over 2.7 kbp and sometimes exceeding 100 kbp in length. Analysis of orthologous gene sets identifies gene families that may be unique to conifers. We further characterize and expand the existing repeat library based on the de novo analysis of the repetitive content, estimated to encompass 82% of the genome.ConclusionsIn addition to its value as a resource for researchers and breeders, the loblolly pine genome sequence and assembly reported here demonstrates a novel approach to sequencing the large and complex genomes of this important group of plants that can now be widely applied.

/pdf/this-provisional-pdf-corresponds-to-the-article-as-it-byjj95ncjz.pdf

This Provisional PDF corresponds to the article as it appeared upon acceptance. Copyedited and fully formatted PDF and full text (HTML) versions will be made available soon. Decoding the massive genome of loblolly pine using haploid DNA and novel assembly strategies

A raison d'être for two distinct pathways in the early steps of plant isoprenoid biosynthesis?

Flavanone 3-hydroxylase (F3H) activity is necessary for the biosynthesis of flavonoids, the main ingredients of Gingko biloba extract. The full-length cDNA and genomic DNA sequences of F3H gene were isolated from G. biloba for the first time. The full-length cDNA of G. biloba F3H gene (designated as GbF3H) contained a 1071 bp open reading frame (ORF) encoding a 357-amino-acid protein with a calculated molecular weight of about 40 kDa and isoelectric point (pI) of 5.57. The genomic DNA analysis showed that GbF3H gene had three exons and two introns. The deduced GbF3H protein showed high identities to other plant F3Hs. The conserved amino acids ligating ferrous iron and residues participating in 2-oxoglutarate binding (R-X-S) were found in GbF3H at the similar positions like other F3Hs. Three-dimensional structure modeling showed that GbF3H had a jerry roll in the enzyme core consisted of β-sheet, a typical structure shared by all 2-oxoglutarate-dependent dioxygenases including F3Hs. Phylogenetic tree analysis revealed that GbF3H shared the same ancestor in evolution with other F3Hs and had a further relationship with other angiosperms species. Southern blot analysis indicated that GbF3H belonged to a multi-gene family. Transcription analysis revealed that GbF3H expressed in stem and leaf with the highest transcription level in leaf. The isolation and characterization of GbF3H gene will be helpful to further study the role of GbF3H gene in the biosynthesis of flavonoids in G. biloba.

Cloning and Characterization of a Flavanone 3-Hydroxylase Gene from Ginkgo biloba

Molecular cloning, characterization and expression of a novel Asr gene from Ginkgo biloba.

2C-methyl-D-erythritol 2, 4-cyclodiphosphate synthase (MECPS, EC: 4.6.1.12) is the fifth enzyme of the non-mevalonate terpenoid pathway for isopentenyl diphosphate biosynthesis and is involved in the methylerythritol phosphate (MEP) pathway for ginkgolide biosynthesis. The full-length mecps cDNA sequence (designated as Gbmecps) was cloned and characterized for the first time from gymnosperm plant species, Ginkgo biloba, using RACE (rapid amplification of cDNA ends) technique. The full-length cDNA of Gbmecps was 874 bp containing a 720 bp open reading frame (ORF) encoding a peptide of 239 amino acids with a calculated molecular mass of 26.03 kDa and an isoelectric point of 8.83. Comparative and bioinformatic analyses revealed that GbMECPS showed extensive homology with MECPSs from other species and contained conserved residues owned by the MECPS protein family. Phylogenetic analysis indicated that GbMECPS was more ancient than other plant MECPSs. Tissue expression pattern analysis indicated that GbMECPS expressed the highest in roots, followed by in leaves, and the lowest in seeds. The color complementation assay indicated that GbMECPS could accelerate the accumulation of beta-carotene. The cloning, characterization and functional analysis of GbMECPS will be helpful to understand more about the role of MECPS involved in the ginkgolides biosynthesis at the molecular level.

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Molecular cloning, characterization and functional analysis of a 2C-methyl- D-erythritol 2, 4-cyclodiphosphate synthase gene from ginkgo biloba.

A new dehydrin ERD10 gene was cloned and characterized from Brassica napus (designated as Bndhn ERD10). The full-length cDNA of Bndhn ERD10 was 1114 bp and contained an open reading frame of 816 bp encoding a protein of 271 amino acid residues. The deduced Bndhn ERD10 protein contained an 8-serine residue domain and two conserved repeats of the characterized lysine-rich-K-segment (KIKEKLPG). Analysis of full-length cDNA and genomic DNA indicated that there were no introns in Bndhn ERD10 gene. The promoter of Bndhn ERD10 was further obtained by genomic walking technology, and analysis of the promoter indicated that the regulation of Bndhn ERD10 was ABA-dependent. Semi-quantitative RT-PCR of different tissues in unstressed B. napus plants indicated that the transcript of Bndhn ERD10 was more abundant in leaf than in stem and root. The expression profiles of Bndhn ERD10 in B. napus seedlings under various stress conditions including cold, salt and ABA were also investigated. Upon cold, salt and ABA stresses,...

A novel ABA-dependent dehydrin ERD10 gene from Brassica napus.

A new CRT binding factor (CBF) gene designated Cbcbf25 was cloned from Capsella bursa-pastoris, a wild grass, by the rapid amplification of cDNA ends (RACE). The full-length cDNA of Cbcbf25 was 898 bp with a 669 bp open reading frame (ORF) encoding a putative DRE/CRT (LTRE)-binding protein of 223 amino acids. The predicted CbCBF25 protein contained a potential nuclear localization signal (NLS) in its N-terminal region followed by an AP2 DNA-binding motif and a possible acidic activation domain in the C-terminal region. Bioinformatic analysis revealed that Cbcbf25 has a high level of similarity with other CBF genes like cbf1, cbf2, and cbf3 from Arabidopsis thaliana, and Bncbf5, Bncbf7, Bncbf16, and Bncbf17 from Brassica napus. A cold acclimation assay showed that Cbcbf25 was expressed immediately after cold triggering, but this expression was transient, suggesting that it concerns cold acclimation. Our study implies that Cbcbf25 is an analogue of other CBF genes and may participate in cold-response, by for example, controlling the expression of cold-regulated genes or increasing the freezing tolerance of plants.

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Zhongxiang Deng

Papers

Cloning and Characterization of a Flavanone 3-Hydroxylase Gene from Ginkgo biloba

Molecular cloning, characterization and expression of a novel Asr gene from Ginkgo biloba.

Molecular cloning, characterization and functional analysis of a 2C-methyl- D-erythritol 2, 4-cyclodiphosphate synthase gene from ginkgo biloba.

A novel ABA-dependent dehydrin ERD10 gene from Brassica napus.

Isolation and molecular characterization of a new CRT binding factor gene from Capsella bursa-pastoris.