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Abstract: Crambe abyssinica (crambe) is a new industrial oil crop that can grow on saline soil and tolerates salty water irrigation. Genetically engineered crambe in which the seed‐oil composition is manipulated for more erucic acid and less polyunsaturated fatty acid (PUFA) would be highly beneficial to industry. In this research, lysophosphatidic acid acyltransferase 2 RNA interference (CaLPAT2‐RNAi) was introduced into the crambe genome to manipulate its oil composition. The result showed in comparison with wild type, CaLPAT2‐RNAi could significantly reduce linoleic and linolenic acid content, simultaneously increasing erucic acid content. Systematic metabolism engineering was then carried out to further study CaLPAT2‐RNAi, combined with the overexpression of Brassica napus fatty acid elongase (BnFAE), Limnanthes douglasii LPAT (LdLPAT), and RNAi of endogenous fatty acid desaturase 2 (CaFAD2‐RNAi). Oil composition analysis on the tranformants' seeds showed that (a) with CaFAD2‐RNAi, PUFA content could be dramatically decreased, in comparison with BnFAE + LdLPAT + CaFAD2‐RNAi, and BnFAE + LdLPAT + CaFAD2‐RNAi + CaLPAT2‐RNAi seeds showed lower linolenic acid content; (b) BnFAE + LdLPAT + CaFAD2‐RNAi + CaLPAT2‐RNAi could increase the erucic acid content in crambe seed oil from less than 66.6% to 71.6%, whereas the highest erucic acid content of BnFAE + LdLPAT + CaFAD2‐RNAi was 79.2%; (c) although the four‐gene combination could not increase the erucic acid content of seed oil to a higher level than the others, it led to increased carbon resource deposited into C22:1 and C18:1 moieties and lower PUFA. Summarily, the present research indicates that suppression of LPAT2 is a new, promising strategy for seed‐oil biosynthesis pathway engineering, which would increase the value of crambe oil. [ABSTRACT FROM AUTHOR]

Water deficit is an environmental factor that constrains crops to express their ecophysiological potential and causes crop yield reduction. Eruca vesicaria has been reported to be one of the most drought-tolerant species in Cruciferae. In this study, polyethylene glycol-simulated drought tolerance was evaluated in one line of Brassica carinata, one line of Brassica napus and 249 Eruca lines based on the principal component analysis (PCA) and unweighted pair-group arithmetic average (UPGMA) cluster analysis. The PCA based on eight drought tolerance indices indicated that the first three components accounted for 85.46% of the total variation, with principal component (PC) 1 accounting for 43.89%, PC2 for 27.85% and PC3 for 13.73% of the total variation. The UPGMA cluster analysis indicated that B. napus cultivar Zhongshuang 9 and Eruca lines could be clustered into five major groups, with group 1 being, in general, drought sensitive, group 2 being slightly–medium drought tolerant, group 3 being drought tolerant, group 4 being highly drought sensitive and group 5 being highly drought tolerant. B. carinata cultivar XB1, as an outstander, showed high drought sensitivity. The UPGMA cluster dendrogram provides a good representation of the similarity matrix (r= 0.68). The drought-tolerant Eruca materials obtained in this study will be valuable for genetic improvement not only in Eruca itself, but also in Brassica crops since they are drought-tolerant lines from a drought-tolerant species. [ABSTRACT FROM PUBLISHER]

We performed a pot culture experiment to evaluate the effects of the heavy metals cadmium and chromium (Cd 1, 10 mg kg−1; Cd 2, 20 mg kg−1; Cd 3, 50 mg kg−1; Cr 1, 100 mg kg−1; Cr 2, 400 mg kg−1; Cr 3, 700 mg kg−1; control, without heavy metals) on the microbial community and carbon utilization diversity. Of 46 bacterial genera (relative abundance ratio of genera greater than 0.3%), 28 and 31 genera were significantly affected by the Cd treatments and Cr treatments, respectively. Soil containing 20 mg kg−1Cd exhibited the highest relative bacterial abundance, bacterial community diversity and carbon utilization diversity. The relative abundance ofLongilineawas significantly higher in the control soil than in heavy metal-treated soils at all sampling stages. The relative abundance ofPseudomonaswas significantly higher in Cd 2 and Cd 3 soils than in the other heavy metal-treated and control soils. The relative abundance of bacteria and carbon source utilization ability were higher at the heading stage than at the maturing stage. Therefore, certain concentrations of heavy metals can improve microbial community structure and carbon utilization diversity. [ABSTRACT FROM PUBLISHER]

Crambe abyssinica is a hexaploid oil crop for industrial applications. An increase of erucic acid (C22:1) and reduction of polyunsaturated fatty acid ( PUFA) contents in crambe oil is a valuable improvement. An increase in oleic acid (C18:1), a reduction in PUFA and possibly an increase in C22:1 can be obtained by down-regulating the expression of fatty acid desaturase2 genes ( Ca FAD2), which code for the enzyme that converts C18:1 into C18:2. We conducted EMS-mutagenesis in crambe, followed by Illumina sequencing, to screen mutations in three expressed Ca FAD2 genes. Two novel analysis strategies were used to detect mutation sites. In the first strategy, mutation detection targeted specific sequence motifs. In the second strategy, every nucleotide position in a Ca FAD2 fragment was tested for the presence of mutations. Seventeen novel mutations were detected in 1100 one-dimensional pools (11 000 individuals) in three expressed Ca FAD2 genes, including non-sense mutations and mis-sense mutations in CaFAD2-C1, - C2 and - C3. The homozygous non-sense mutants for Ca FAD2-C3 resulted in a 25% higher content of C18:1 and 25% lower content of PUFA compared to the wild type. The mis-sense mutations only led to small changes in oil composition. Concluding, targeted mutation detection using NGS in a polyploid was successfully applied and it was found that a non-sense mutation in even a single Ca FAD2 gene can lead to changes in crambe oil composition. Stacking the mutations in different Ca FAD2 may gain additional changes in C18:1 and PUFA contents. [ABSTRACT FROM AUTHOR]

Crambe abyssinica is a hexaploid oil crop for industrial applications. An increase of erucic acid (C22:1) and reduction of polyunsaturated fatty acid (PUFA) contents in crambe oil is a valuable improvement. An increase in oleic acid (C18:1), a reduction in PUFA and possibly an increase in C22:1 can be obtained by down-regulating the expression of fatty acid desaturase2 genes (CaFAD2), which code for the enzyme that converts C18:1 into C18:2. We conducted EMS-mutagenesis in crambe, followed by Illumina sequencing, to screen mutations in three expressed CaFAD2 genes. Two novel analysis strategies were used to detect mutation sites. In the first strategy, mutation detection targeted specific sequence motifs. In the second strategy, every nucleotide position in a CaFAD2 fragment was tested for the presence of mutations. Seventeen novel mutations were detected in 1100 one-dimensional pools (11 000 individuals) in three expressed CaFAD2 genes, including non-sense mutations and mis-sense mutations in CaFAD2-C1, -C2 and -C3. The homozygous non-sense mutants for CaFAD2-C3 resulted in a 25% higher content of C18:1 and 25% lower content of PUFA compared to the wild type. The mis-sense mutations only led to small changes in oil composition. Concluding, targeted mutation detection using NGS in a polyploid was successfully applied and it was found that a non-sense mutation in even a single CaFAD2 gene can lead to changes in crambe oil composition. Stacking the mutations in different CaFAD2 may gain additional changes in C18:1 and PUFA contents., (© 2014 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.)