Although abscisic acid (ABA) is an important hormone that regulates seed dormancy, stomatal closure, plant development, as well as responses to environmental stimuli, the physiological mechanisms of ABA response to multiple stress in rice remain poorly understood. In the ABA biosynthetic pathway, 9-cis-epoxycarotenoid dioxygenase (NCED) is the key rate-limiting enzyme. Here, we report important functions of OsNCED3 in multi-abiotic stress tolerance in rice. The OsNCED3 is constitutively expressed in various tissues under normal condition, Its expression is highly induced by NaCl, PEG, and H2O2 stress, suggesting the roles for OsNCED3 in response to the multi-abiotic stress tolerance in rice. Compared with wild-type plants, nced3 mutants had earlier seed germination, longer post-germination seedling growth, increased sensitivity to water stress and H2O2 stress and increased stomata aperture under water stress and delayed leaf senescence. Further analysis found that nced3 mutants contained lower ABA content compared with wild-type plants, overexpression of OsNCED3 in transgenic plants could enhance water stress tolerance, promote leaf senescence and increase ABA content. We conclude that OsNCED3 mediates seed dormancy, plant growth, abiotic stress tolerance, and leaf senescence by regulating ABA biosynthesis in rice; and may provide a new strategy for improving the quality of crop.

Database: The C. gracea KCS sequence information has been submitted to the EMBL ⁄ GenBank under accession number: bankit1110928.Keywords: 3-keto-acyl-CoA synthase, Cardamine graeca, Nervonic acid, Brassica carinata, Brassica napus, Arabidopsis thaliana, Health and Industry. SummaryNervonic acid 24:1 D15 (cis-tetracos-15-enoic acid) is a very long-chain monounsaturated fatty acid and exists in nature as an elongation product of oleic acid.There is an increasing interest in production of high nervonic acid oils for pharmaceutical, nutraceutical and industrial applications. Using a polymerase chain reaction approach, we have isolated a gene from Cardamine graeca L., which encodes a 3-ketoacyl-CoA synthase (KCS), the first component of the elongation complex involved in synthesis of nervonic acid. Expression of the Cardamine KCS in yeast resulted in biosynthesis of nervonic acid, which is not normally present in yeast cells.We transformed Arabidopsis and Brassica carinata with the Cardamine KCS under the control of the seed-specific promoter, napin. The T 3 generations of transgenic Arabidopsis and B. carinata plants expressing the Cardamine KCS showed that seed-specific expression resulted in relatively large comparative increases in nervonic acid proportions in Arabidopsis seed oil, and 15-fold increase in nervonic acid proportions in B. carinata seed oil. The highest nervonic acid level in transgenic B. carinata lines reached 44%, with only 6% of residual erucic acid. In contrast, similar transgenic expression of the Cardamine KCS in high erucic B. napus resulted in 30% nervonic acid but with 20% residual erucic acid. Experiments using the Lunaria KCS gene gave results similar to the latter. In both cases, the erucic acid content is too high for human or animal consumption. Thus, the Cardamine KCS: B. carinata high nervonic ⁄ highly reduced erucic transgenic seed oils will be the most suitable for testing in pharmaceutical ⁄ nutraceutical applications to improve human and animal health.

Nervonic acid is a Very Long-Chain Monounsaturated Fatty Acid (VLCMFA), 24:1 Delta15 (cis-tetracos-15-enoic acid) found in the seed oils of Lunaria annua, borage, hemp, Acer (Purpleblow maple) and Tropaeolum speciosum (Flame flower). However, of these, only the "money plant" (Lunaria annua L.) has been studied and grown sparingly for future development as a niche crop and the outlook has been disappointing. Therefore, our goal was to isolate and characterize strategic new genes for high nervonic acid production in Brassica oilseed crops. To this end, we have isolated a VLCMFA-utilizing 3-Keto-Acyl-CoA Synthase (KCS; fatty acid elongase; EC 2.3.1.86) gene from Lunaria annua and functionally expressed it in yeast, with the recombinant KCS protein able to catalyze the synthesis of several VLCMFAs, including nervonic acid. Seed-specific expression of the Lunaria KCS in Arabidopsis resulted in a 30-fold increase in nervonic acid proportions in seed oils, compared to the very low quantities found in the wild-type. Similar transgenic experiments using B. carinata as the host resulted in a 7-10 fold increase in seed oil nervonic acid proportions. KCS enzyme activity assays indicated that upon using (14)C-22:1-CoA as substrate, the KCS activity from developing seeds of transgenic B. carinata was 20-30-fold higher than the low erucoyl-elongation activity exhibited by wild type control plants. There was a very good correlation between the Lun KCS transcript intensity and the resultant 22:1-CoA KCS activity in developing seed. The highest nervonic acid level in transgenic B. carinata expressing the Lunaria KCS reached 30%, compared to 2.8% in wild type plant. In addition, the erucic acid proportions in these transgenic lines were considerably lower than that found in native Lunaria oil. These results show the functional utility of the Lunaria KCS in engineering new sources of high nervonate/reduced erucic oils in the Brassicaceae.

Brassica rapa is the most widely distributed and has the longest history of domestication of the agricultural Brassica species. Molecular genetic diversity, based on 51 simple sequence repeat primer pairs and 715 alleles at polymorphic loci, was used to predict the center of origin and centers of diversity in a global collection of 173 B. rapa accessions. The accessions were separated into 3 molecular genetic groups based on STRUCTURE analysis-group 1 from the classical Old World (Europe and west Asia-north Africa), group 2 from east Asia, and group 3 from east, central, south, and southeast Asia. Accessions classified as "wild" (B. rapa var. sylvestris) were found only in group 1 and this group had the highest number and richness of private alleles. Each group included a diverse range of agricultural morphotypes (oilseed, root, or leafy vegetable types), flowering habit (winter, semi-winter, or spring type), self-compatibility or incompatibility, and seed color. The Old World and east, south, and central Asia were distinct subpopulations based on analysis of shared unique alleles. This study supports the theory that the classical Old World is the center of origin of B. rapa, with centers of diversity in east Asia and along ancient trade routes in Asia, with recent migration to the New World.

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Oilseed Brassica species are vulnerable to heat and drought stress, especially in the early reproductive stage. We evaluated plant imaging of whole plant and flower tissue, leaf stomatal conductance, leaf and bud temperature, photochemical reflectance index, quantum yield of photosynthesis, and leaf gas exchange for their suitability to detect tolerance to heat (H) and/or drought (D) stress treatments in 12 Brassica genotypes (G). A replicated factorial experiment was set up with 7 d of stress treatment from the beginning of anthesis with various levels of three factors H, D, and G. Most phenomics tools detected plant stress as indicated by significant main effects of H, D, and H×D. Whole plant volume was highly correlated with fresh weight changes, suggesting that whole plant imaging may be a useful surrogate for fresh weight in future studies. Vcmax, the maximum carboxylation rate of photosynthesis, increased rapidly on day 1 in H and H+D treatments, and there were significant interactions of G×H and G×D. Vcmax of genotypes on day 1 in H and H+D treatments was positively correlated with their harvested seed yield. Vcmax on day 1 and day 3 were clustered with seed yield in H and H+D treatments as shown in the heatmaps of genotypic correlations. TPU, the rate of triose phosphate use, also showed significant positive genotypic correlations with seed yield in H+D treatments. Flower volume showed significant interactions of G×H and G×D on day 7, and flower volume of genotypes on day 7 in H was positively correlated with their harvested seed yield. There were few interactions of G×H or G×D for leaf stomatal conductance, leaf and bud temperature, photochemical reflectance index, and quantum yield of photosynthesis. Vcmax, TPU, and volume of flowers are potential nondestructive phenomic traits for heat or combined heat and drought stress tolerance screening in Brassica germplasm.

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