Vincent Vadez

Transgenic approaches for abiotic stress tolerance in plants: retrospect and prospects.

Abiotic stresses including drought are serious threats to the sustainability of crop yields accounting for more crop productivity losses than any other factor in rainfed agriculture. Success in breeding for better adapted varieties to abiotic stresses depend upon the concerted efforts by various research domains including plant and cell physiology, molecular biology, genetics, and breeding. Use of modern molecular biology tools for elucidating the control mechanisms of abiotic stress tolerance, and for engineering stress tolerant crops is based on the expression of specific stress-related genes. Hence, genetic engineering for developing stress tolerant plants, based on the introgression of genes that are known to be involved in stress response and putative tolerance, might prove to be a faster track towards improving crop varieties. Far beyond the initial attempts to insert “single-action” genes, engineering of the regulatory machinery involving transcription factors has emerged as a new tool now for controlling the expression of many stress-responsive genes. Nevertheless, the task of generating transgenic cultivars is not only limited to the success in the transformation process, but also proper incorporation of the stress tolerance. Evaluation of the transgenic plants under stress conditions, and understanding the physiological effect of the inserted genes at the whole plant level remain as major challenges to overcome. This review focuses on the recent progress in using transgenic technology for the improvement of abiotic stress tolerance in plants. This includes discussion on the evaluation of abiotic stress response and the protocols for testing the transgenic plants for their tolerance under close-to-field conditions.

Genetic variability of drought-avoidance root traits in the mini-core germplasm collection of chickpea (Cicer arietinum L.).

Extensive and deep root systems have been recognized as one of the most important traits for improving chickpea (Cicer arietinum L.) productivity under progressively receding soil moisture conditions. However, available information on the range of variation for root traits is still limited. Genetic variability for the root traits was investigated using a cylinder culture system during two consecutive growth seasons in the mini-core germplasm collection of ICRISAT plus several wild relatives of chickpea. The largest genetic variability was observed at 35 days after sowing for root length density (RLD) (heritability, h2 = 0.51 and 0.54) across seasons, and followed by the ratio of plant dry weight to root length density with h2 of 0.37 and 0.50 for first and second season, respectively. The root growth of chickpea wild relatives was relatively poor compared to C. arietinum, except in case of C. reticulatum. An outstanding genotype, ICC 8261, which had the largest RLD and one of the deepest root system, was identified in chickpea mini-core germplasm collection. The accession ICC 4958 which was previously characterized as a source for drought avoidance in chickpea was confirmed as one with the most prolific and deep root system, although many superior accessions were also identified. The chickpea landraces collected from the Mediterranean and the west Asian region showed a significantly larger RLD than those from the south Asian region. In addition, the landraces originating from central Asia (former Soviet Union), characterized by arid agro-climatic conditions, also showed relatively larger RLD. As these regions are under-represented in the chickpea collection, they might be interesting areas for further germplasm exploration to identify new landraces with large RLD. The information on the genetic variability of chickpea root traits provides valuable baseline knowledge for further progress on the selection and breeding for drought avoidance root traits in chickpea.

Stress-inducible expression of At DREB1A in transgenic peanut (Arachis hypogaea L.) increases transpiration efficiency under water-limiting conditions

Water deficit is the major abiotic constraint affecting crop productivity in peanut (Arachis hypogaea L.). Water use efficiency under drought conditions is thought to be one of the most promising traits to improve and stabilize crop yields under intermittent water deficit. A transcription factor DREB1A from Arabidopsis thaliana, driven by the stress inducible promoter from the rd29A gene, was introduced in a drought-sensitive peanut cultivar JL 24 through Agrobacterium tumefaciens-mediated gene transfer. The stress inducible expression of DREB1A in these transgenic plants did not result in growth retardation or visible phenotypic alterations. T3 progeny of fourteen transgenic events were exposed to progressive soil drying in pot culture. The soil moisture threshold where their transpiration rate begins to decline relative to control well-watered (WW) plants and the number of days needed to deplete the soil water was used to rank the genotypes using the average linkage cluster analysis. Five diverse events were selected from the different clusters and further tested. All the selected transgenic events were able to maintain a transpiration rate equivalent to the WW control in soils dry enough to reduce transpiration rate in wild type JL 24. All transgenic events except one achieved higher transpiration efficiency (TE) under WW conditions and this appeared to be explained by a lower stomatal conductance. Under water limiting conditions, one of the selected transgenic events showed 40% higher TE than the untransformed control.

A comprehensive resource of drought- and salinity- responsive ESTs for gene discovery and marker development in chickpea (Cicer arietinum L.)

BackgroundChickpea (Cicer arietinum L.), an important grain legume crop of the world is seriously challenged by terminal drought and salinity stresses. However, very limited number of molecular markers and candidate genes are available for undertaking molecular breeding in chickpea to tackle these stresses. This study reports generation and analysis of comprehensive resource of drought- and salinity-responsive expressed sequence tags (ESTs) and gene-based markers.ResultsA total of 20,162 (18,435 high quality) drought- and salinity- responsive ESTs were generated from ten different root tissue cDNA libraries of chickpea. Sequence editing, clustering and assembly analysis resulted in 6,404 unigenes (1,590 contigs and 4,814 singletons). Functional annotation of unigenes based on BLASTX analysis showed that 46.3% (2,965) had significant similarity (≤1E-05) to sequences in the non-redundant UniProt database. BLASTN analysis of unique sequences with ESTs of four legume species (Medicago, Lotus, soybean and groundnut) and three model plant species (rice, Arabidopsis and poplar) provided insights on conserved genes across legumes as well as novel transcripts for chickpea. Of 2,965 (46.3%) significant unigenes, only 2,071 (32.3%) unigenes could be functionally categorised according to Gene Ontology (GO) descriptions. A total of 2,029 sequences containing 3,728 simple sequence repeats (SSRs) were identified and 177 new EST-SSR markers were developed. Experimental validation of a set of 77 SSR markers on 24 genotypes revealed 230 alleles with an average of 4.6 alleles per marker and average polymorphism information content (PIC) value of 0.43. Besides SSR markers, 21,405 high confidence single nucleotide polymorphisms (SNPs) in 742 contigs (with ≥ 5 ESTs) were also identified. Recognition sites for restriction enzymes were identified for 7,884 SNPs in 240 contigs. Hierarchical clustering of 105 selected contigs provided clues about stress- responsive candidate genes and their expression profile showed predominance in specific stress-challenged libraries.ConclusionGenerated set of chickpea ESTs serves as a resource of high quality transcripts for gene discovery and development of functional markers associated with abiotic stress tolerance that will be helpful to facilitate chickpea breeding. Mapping of gene-based markers in chickpea will also add more anchoring points to align genomes of chickpea and other legume species.

Terminal drought-tolerant pearl millet [Pennisetum glaucum (L.) R. Br.] have high leaf ABA and limit transpiration at high vapour pressure deficit

It was previously shown that pearl millet genotypes carrying a terminal drought tolerance quantitative trait locus (QTL) had a lower transpiration rate (Tr; g cm−2 d−1) under well-watered conditions than sensitive lines. Here experiments were carried out to test whether this relates to leaf abscisic acid (ABA) and Tr concentration at high vapour pressure deficit (VPD), and whether that leads to transpiration efficiency (TE) differences. These traits were measured in tolerant/sensitive pearl millet genotypes, including near-isogenic lines introgressed with a terminal drought tolerance QTL (NIL-QTLs). Most genotypic differences were found under well-watered conditions. ABA levels under well-watered conditions were higher in tolerant genotypes, including NIL-QTLs, than in sensitive genotypes, and ABA did not increase under water stress. Well-watered Tr was lower in tolerant than in sensitive genotypes at all VPD levels. Except for one line, Tr slowed down in tolerant lines above a breakpoint at 1.40–1.90 kPa, with the slope decreasing >50%, whereas sensitive lines showed no change in that Tr response across the whole VPD range. It is concluded that two water-saving (avoidance) mechanisms may operate under well-watered conditions in tolerant pearl millet: (i) a low Tr even at low VPD conditions, which may relate to leaf ABA; and (ii) a sensitivity to higher VPD that further restricts Tr, which suggests the involvement of hydraulic signals. Both traits, which did not lead to TE differences, could contribute to absolute water saving seen in part due to dry weight increase differences. This water saved would become critical for grain filling and deserves consideration in the breeding of terminal drought-tolerant lines.

Constitutive water-conserving mechanisms are correlated with the terminal drought tolerance of pearl millet [Pennisetum glaucum (L.) R. Br.]

Pearl millet, a key staple crop of the semi-arid tropics, is mostly grown in water-limited conditions, and improving its performance depends on how genotypes manage limited water resources. This study investigates whether the control of water loss under non-limiting water conditions is involved in the terminal drought tolerance of pearl millet. Two pairs of tolerant×sensitive pearl millet genotypes, PRLT 2/89-33–H77/833-2 and 863B-P2–ICMB 841-P3, and near-isogenic lines (NILs), introgressed with a terminal drought tolerance quantitative trait locus (QTL) from the donor parent PRLT 2/89-33 into H77/833-2 (NILs-QTL), were tested. Upon exposure to water deficit, transpiration began to decline at lower fractions of transpirable soil water (FTSW) in tolerant than in sensitive genotypes, and NILs-QTL followed the pattern of the tolerant parents. The transpiration rate (Tr, in g water loss cm−2 d−1) under well-watered conditions was lower in tolerant than in sensitive parental genotypes, and the Tr of NILs-QTL followed the pattern of the tolerant parents. In addition, Tr measured in detached leaves (g water loss cm−2 h−1) from field-grown plants of the parental lines showed lower Tr values in tolerant parents. Defoliation led to an increase in Tr that was higher in sensitive than in tolerant genotypes. The differences in Tr between genotypes was not related to the stomatal density. These results demonstrate that constitutive traits controlling leaf water loss under well-watered conditions correlate with the terminal drought tolerance of pearl millet. Such traits may lead to more water being available for grain filling under terminal drought.

A conservative pattern of water use, rather than deep or profuse rooting, is critical for the terminal drought tolerance of chickpea

Chickpea is mostly grown on stored soil moisture, and deep/profuse rooting has been hypothesized for almost three decades to be critical for improving chickpea tolerance to terminal drought. However, temporal patterns of water use that leave water available for reproduction and grain filling could be equally critical. Therefore, variation in water use pattern and root depth/density were measured, and their relationships to yield tested under fully irrigated and terminal drought stress, using lysimeters that provided soil volumes equivalent to field conditions. Twenty chickpea genotypes having similar plant phenology but contrasting for a field-derived terminal drought-tolerance index based on yield were used. The pattern of water extraction clearly discriminated tolerant and sensitive genotypes. Tolerant genotypes had a lower water uptake and a lower index of stomatal conductance at the vegetative stage than sensitive ones, while tolerant genotypes extracted more water than sensitive genotypes after flowering. The magnitude of the variation in root growth components (depth, length density, RLD, dry weight, RDW) did not distinguish tolerant from sensitive genotypes. The seed yield was not significantly correlated with the root length density (RLD) in any soil layers, whereas seed yield was both negatively related to water uptake between 23–38 DAS, and positively related to water uptake between 48–61 DAS. Under these conditions of terminal drought, the most critical component of tolerance in chickpea was the conservative use of water early in the cropping cycle, explained partly by a lower canopy conductance, which resulted in more water available in the soil profile during reproduction leading to higher reproductive success.

Physiological traits for crop yield improvement in low N and P environments

Nitrogen and phosphorus are recognized as essential elements in crop production, but the full extent of the requirement for these elements in the physiological processes leading to crop growth seems not to be always fully appreciated. Virtually all the biochemical compounds in plants that support development and growth contain N and/or P. Deficiencies in either element lead to a lost ability for plant growth such that there is a quantitative relationship between crop yield and accumulation by plants of each of these elements. Few options appear to exist to greatly diminish the requirement for either element in crop growth and the formation of seed yield. Consequently, crop yields cannot be increased without increased acquisition of N and P by plants. If the soil environment does not offer these elements, then crop yield will necessarily be restricted. While little opportunity exists to increase N recovery under low nutrient environments, several options can be investigated for increasing P accumulation by the crop. Ultimately, however, the rigid limitation on yields of inadequate N means that without external supplies of N for the cropping system, biological fixation of N must be enhanced to increase N input. In particular, it appears that considerable research needs to be focused on whole-plant processes in legumes that lead to enhanced symbiotic N fixation. A critical aspect of increased legume production will be improved management of P to allow legumes to achieve high N fixation rates and yields.

Cultural, Practical, and Economic Value of Wild Plants: a Quantitative Study in the Bolivian Amazon

Researchers have developed several indices to estimate the significance of plant species for humans. We build on previous methods in ethnobotany and anthropology to develop a new way to value plant species along three dimensions: cultural, practical, and economic. We used interview and observational data on the use of wild plants by the Tsimane’, a foraging-horticultural society in the Bolivian Amazon. We calculated the cultural, practical, economic, and total values of 114 plant species from 46 families. We found a low correlation between the practical and the cultural values of species: some species rarely used were frequently mentioned in interviews, whereas some species frequently used were rarely mentioned in interviews. Indices of cultural, practical, and economic value measure different dimensions of the importance of plant species to society. The combination of the three indices offers a more comprehensive valuation of the significance of plants for humans than the use of only one index.

Market Economy and the Loss of Folk Knowledge of Plant Uses: Estimates from the Tsimane' of the Bolivian Amazon

victoria reyes -garc ı́a , v incent vadez , el izabeth byron, l il ian apaza, will iam r. leonard, eddy perez , and david wilkie Sustainable International Development Program, Heller School for Social Policy and Management, Brandeis University, Waltham, MA 02454-9110, U.S.A. (vreyes@brandeis.edu) (Reyes-Garcı́a and Vadez)/International Food Policy Research Institute, Washington, DC 20006-1002, U.S.A. (Byron)/ Protección del Medio Ambiente Tarija, Calle Alejandro del Carpio N E-0659, Casilla N 59, Bolivia (Apaza)/Department of Anthropology, Northwestern University, Evanston, IL 60208, U.S.A. (Leonard)/ Fundación para el Desarrollo de la Ecologia, Estación Biológica Tunquini, Bolivia (Pérez)/Wildlife Conservation Society, 18 Clark Lane, Waltham, MA 02451-1823, U.S.A. (Wilkie). 10 ii 05