Effects of α-Fe2O3 nanoparticles and biochar on plant growth and fruit quality of muskmelon under cadmium stress.

Cadmium pollution in farmland has become a global environmental problem, threatening ecological security and human health. Biochar is effective in remediation of soil pollution. However, high concentrations of biochar can inhibit plant growth, and low concentrations of biochar have limited mitigation effect on cadmium toxicity. Therefore, the combination of low-concentration biochar and other amendments is a promising approach to alleviate cadmium toxicity in plants and improve the safety of edible parts. In this study, muskmelon was selected as the research object, and different concentrations of α-Fe2O3 nanoparticles were used alone or combined with biochar to explore the effects of different treatments on muskmelon plants in cadmium-contaminated soil. The results showed that the combined application of 250 mg/kg α-Fe2O3 nanoparticles and biochar had a good effect on the repair of cadmium toxicity in muskmelon plants. Compared with cadmium treatment, its application increased plant height by 32.53%, cadmium transport factor from root to stem decreased by 32.95%, chlorophyll content of muskmelon plants increased by 14.27%, and cadmium content in muskmelon flesh decreased by 18.83%. Moreover, after plant harvest, soil available cadmium content in 250 mg/kg α-Fe2O3 nanoparticles and biochar combined treatment decreased by 31.18% compared with cadmium treatment. The results of this study provide an effective reference for the composite application of different exogenous amendments and a feasible idea for soil heavy metal remediation and mitigation of cadmium pollution in farmland.

Transposon insertions within alleles of BnaFT.A2 are associated with seasonal crop type in rapeseed.

KEY MESSAGE: We identified two new transposon insertions within the promoter of BnaFT.A2 in addition to an existing 288 bp MITE within the second intron. Each insertion event corresponds to a distinct BnaFT.A2 haplotype and is closely associated with established crop seasonal ecotypes. Florigen, encoded by FLOWERING LOCUS T (FT), plays key roles not only as a flowering hormone, but also a universal growth factor affecting several aspects of plant architecture. In rapeseed, BnaFT.A2 has been revealed as one of the major loci associated with flowering time and different ecotypes. However, it is unclear how allelic variations of BnaFT.A2 affect its function in flowering time regulation and beyond. In this study, we confirmed an existing 288 bp miniature inverted-repeat transposable element (MITE) insertion within the second intron and identified two new insertions within the promoter of BnaFT.A2-a 3971 bp CACTA and a 1079 bp Helitron. Each insertion event corresponds to a distinct BnaFT.A2 haplotype and is closely associated with established crop seasonal ecotypes. These alleles have similar tissue-specific expression patterns but discrete transcriptional patterns tightly associated with rapeseed flowering time and ecotype. RNAi lines and mutants of BnaFT.A2 flowered significantly later than controls. Differentially expressed genes (DEGs), identified in transcriptomic profiling of seedling leaves from two loss-of-function mutants (Bnaft.a2-L1 and Bnaft.a2-L2) compared with controls, indicated significant enrichment for hormone metabolic genes and roles related to plant cell wall synthesis and photosynthesis. Plants with loss-of-function BnaFT.A2 had smaller leaves and lower net photosynthetic rate compared to controls. These findings not only further clarify the genetic basis of flowering time variation and ecotype formation in B. napus, but also provide an additional toolbox for genetic improvement of seasonal adaptation and production.

Effects of Cadmium on Physiochemistry and Bioactive Substances of Muskmelon (Cucumis melo L.).

Muskmelon pedicel is the fruit stalk of muskmelon and one of the traditional Chinese medicines, which can be used to treat jaundice, diabetes and neuropathy. However, in recent years, agricultural soil heavy metal cadmium (Cd) pollution has become serious, coupled with the imperfect sales management of herbal medicine, increasing the potential health risk of contaminated herbal medicine in the human body. In this paper, the comprehensive quality of contaminated muskmelon was tested. The results showed that Cd stress significantly inhibited the growth of muskmelon plants, reduced the anthocyanin and chlorophyll contents, and increased the fruit size and sweetness of muskmelon. In addition, heavy metal Cd can also cause oxidative stress in plants, resulting in a series of changes in antioxidant enzyme activities. In the experimental group, the content of polyphenols and saponins increased by 27.02% and 23.92%, respectively, after high-concentration Cd treatment, which may be a mechanism of plant resistance to stress. This paper reveals that the content of bioactive substances in Chinese herbal medicine is high, but the harm in heavy metals cannot be underestimated, which should be paid attention to by relevant departments.

Physiological of biochar and α-Fe2O3 nanoparticles as amendments of Cd accumulation and toxicity toward muskmelon grown in pots.

BACKGROUND: Due to the severe cadmium (Cd) pollution of farmland soil, effective measures need to be taken to reduce the Cd content in agricultural products. In this study, we added α-Fe2O3 nanoparticles (NPs) and biochar into Cd-contaminated soil to investigate physiological responses of muskmelon in the whole life cycle. RESULTS: The results showed that Cd caused adverse impacts on muskmelon (Cucumis melo) plants. For instance, the chlorophyll of muskmelon leaves in the Cd alone treatment was reduced by 8.07-32.34% in the four periods, relative to the control. The treatments with single amendment, α-Fe2O3 NPs or 1% biochar or 5% biochar, significantly reduced the soil available Cd content, but the co-exposure treatments (α-Fe2O3 NPs and biochar) had no impact on the soil available Cd content. All treatments could reduce the Cd content by 47.64-74.60% and increase the Fe content by 15.15-95.27% in fruits as compared to the Cd alone treatment. The KEGG enrichment results of different genes in different treatments indicated that single treatments could regulate genes related to anthocyanin biosynthesis, glutathione metabolism and MAPK signal transduction pathways to reduce the Cd toxicity. CONCLUSIONS: Overall the combination of biochar and α-Fe2O3 NPs can alleviate Cd toxicity in muskmelon. The present study could provide new insights into Cd remediation in soil using α-Fe2O3 NPs and biochar as amendments.

Genome-wide identification of flowering time genes in cucurbit plants and revealed a gene ClGA2/KS associate with adaption and flowering of watermelon.

Watermelon (Citrullus lanatus) is one of the major cucurbit crop that cultivated all over the world. Adaptability and flowering time are important agronomic characteristics that influence the quality and yield of watermelon, however, the molecular basis underlying these traits were still unclear. In this study, we identified 166, 182, 178, and 279 flowering genes in watermelon, melon, cucumber and pumpkin, respectively, and found that a lot of genes in the photoperiodic, autonomous, and vernalization pathways were absence in the four cucurbits. A higher ratio of flowering time genes was identified in the hormone pathway in cucurbits than in Arabidopsis, and a higher average ka/ks value of hormone pathway genes than the photoperiodic and vernalization pathway genes was identified in watermelon. Moreover, a gene ClGA2/KS (Cla005482) were found to associated with ecotype differentiation, flowering time, and whole growth period in watermelon. This study added knowledge to the molecular basis of flowering time regulation in cucurbits, and the molecule marker of ClGA2/KS gene may facilitate the breeding progress for selecting watermelon varieties with superior adaption and flowering time.

Sequence variation and functional analysis of a FRIGIDA orthologue (BnaA3.FRI) in Brassica napus.

BACKGROUND: Allelic variation at the FRIGIDA (FRI) locus is a major contributor to natural variation of flowering time and vernalization requirement in Arabidopsis thaliana. Dominant FRI inhibits flowering by activating the expression of the MADS box transcriptional repressor FLOWERING LOCUS C (FLC), which represses flowering prior to vernalization. Four FRI orthologues had been identified in the domesticated amphidiploid Brassica napus. Linkage and association studies had revealed that one of the FRI orthologues, BnaA3.FRI, contributes to flowering time variation and crop type differentiation. RESULTS: Sequence analyses indicated that three out of the four BnaFRI paralogues, BnaA3.FRI, BnaA10.FRI and BnaC3.FRI, contained a large number of polymorphic sites. Haplotype analysis in a panel of 174 B. napus accessions using PCR markers showed that all the three paralogues had a biased distribution of haplotypes in winter type oilseed rape (P < 0.01). Association analysis indicated that only BnaA3.FRI contributes to flowering time variation in B. napus. In addition, transgenic functional complementation demonstrated that mutations in the coding sequence of BnaA3.FRI lead to weak alleles, and subsequently to flowering time variation. CONCLUSION: This study for the first time provides a molecular basis for flowering time control by BnaA3.FRI in B. napus, and will facilitate predictive oilseed rape breeding to select varieties with favorable flowering time and better adaption to latitude and seasonal shifts due to changing climate.

Generation and characterization of tribenuron-methyl herbicide-resistant rapeseed (Brasscia napus) for hybrid seed production using chemically induced male sterility.

KEY MESSAGE: Identification and molecular analysis of four tribenuron-methyl resistant mutants in Brassica napus , which would be very useful in hybrid production using a Chemically induced male sterility system. Chemically induced male sterility (CIMS) systems dependent on chemical hybridization agents (CHAs) like tribenuron-methyl (TBM) represent an important approach for practical utilization of heterosis in rapeseed. However, when spraying the female parents with TBM to induce male sterility the male parents must be protected with a shield to avoid injury to the stamens, which would otherwise complicate the seed production protocol and increase the cost of hybrid seed production. Here we report the first proposed application of a herbicide-resistant cultivar in hybrid production, using a CIMS system based on identifying four TBM-resistant mutants in Brassica napus. Genetic analysis indicated that the TBM resistance was controlled by a single dominant nuclear gene. An in vitro enzyme activity assay for acetohydroxyacid synthase (AHAS) suggested that the herbicide resistance is caused by a gain-of-function mutation in a copy of AHAS genes. Comparative sequencing of the mutants and wild type BnaA.AHAS.a coding sequences identified a C-to-T transition at either position 535 or 536 from the translation start site, which resulted in a substitution of proline with serine or leucine at position 197 according to the Arabidopsis thaliana protein sequence. An allele-specific dCAPS marker developed from the C536T variation co-segregated with the herbicide resistance. Transgenic A. thaliana plants expressing BnaA.ahas3.a conferred herbicide resistance, which confirmed that the P197 substitution in BnaA.AHAS.a was responsible for the herbicide resistance. Moreover, the TBM-resistant lines maintain normal male fertility under TBM treatment and can be of practical value in hybrid seed production using CIMS.

Genetic mapping of a single nuclear locus determines the white flesh color in watermelon (Citrullus lanatus L.).

Introduction: Flesh color is an important trait in watermelon (Citrullus lanatus L.). Several flesh color genes have been identified in watermelon; however, the inheritance of and the molecular basis underlying the white flesh trait remain largely unknown. Methods: In this study, segregation populations were constructed by crossing the canary yellow flesh line HSH-F with the white flesh line Sanbai to fine-map the white flesh gene in watermelon. Results: Genetic analysis indicated that the white flesh trait is controlled by a single recessive locus, termed Clwf2. Map-based cloning delimited the Clwf2 locus to a 132.3-kb region on chromosome 6. The candidate region contains 13 putative genes, and four of them-Cla97C06G121860, Cla97C06G121880, Cla97C06G121890, and Cla97C06G121900-were significantly downregulated in the white flesh compared to the canary yellow flesh watermelon fruits. The Cla97C06G121890 gene, which encodes a tetratricopeptide repeat protein, showed almost no expression in the white flesh fruit before maturity, whereas it had a very high expression in the canary yellow flesh fruit at 18 days after pollination. Transmission electron microscopy revealed rounded and regularly shaped chromoplasts in both the canary yellow and white flesh fruits. Further quantitative real-time PCR analysis showed that the expression levels of several key plastid division genes and almost the entire carotenoid biosynthesis pathway genes were downregulated in the white flesh compared to the canary yellow flesh fruits. Discussion: This study suggests that the proliferation inhibition of chromoplasts and downregulation of the CBP genes block the accumulation of carotenoids in watermelon and lead to white flesh. These findings advance and extend the understanding of the molecular mechanisms underlying white flesh trait formation and carotenoid biosynthesis in watermelon.

A study on the mixture repairing effect of biochar and nano iron oxide on toxicity of Cd toward muskmelon.

Soil contamination with cadmium (Cd) has become a serious problem, adversely affecting food safety and human health. Effective methods are urgently needed to alleviate toxicity of Cd in plants. In this study, a nine-week continuous pot experiments was conducted to explore the effectiveness of the different nano iron oxide (α-Fe2O3, γ-Fe2O3, Fe3O4) alone and combined with biochar in muskmelon grown on a Cd-contaminated soil. The antioxidant system, chlorophyll, soluble protein, other physiological indexes of muskmelon leaves and the distribution of Cd in matrix soil, leaves and fruit were detected. The results showed that Cd was readily absorbed by plants and caused oxidative stress on plants, while biochar, α-Fe2O3 nanoparticles (NPs) and their mixture group (BFe1 group) could significantly improve it. Specifically, the three treatments reduced the Cd content of the fruit by 19.51-78.86%, reduced the Cd content of leaves by 15.44-36.23% and 22.36-31.77% in weeks 3 and 5, respectively. For the activity of enzymes, three treatments decreased superoxide dismutase (SOD) activity and catalase (CAT) activity by 3.41-38.57% and 24.27-30.33% in week 7, respectively. So BFe1 group application immobilized Cd in soil and reduced Cd partitioning in the aboveground tissues. Overall the combination of biochar and α-Fe2O3 NPs can alleviate Cd toxicity in muskmelon and can protect human beings from Cd exposure.

Characterization and site-directed mutagenesis of a novel class II 5-enopyruvylshikimate-3-phosphate (EPSP) synthase from the deep-sea bacterium Alcanivorax sp. L27.

The 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) is a key enzyme in the aromatic amino acid biosynthetic pathway in microorganisms and plants, which catalyzes the formation of 5-enolpyruvylshikimate-3-phosphate (EPSP) from shikimate-3-phosphate (S3P) and phosphoenolpyruvate (PEP). In this study, a novel AroA-encoding gene was identified from the deep sea bacterium Alcanivorax sp. L27 through screening the genomic library and termed as AroAA.sp. A phylogenetic analysis revealed that AroAA.sp (1317 bp and 438 amino acids) is a class II AroA. This enzyme exhibited considerable activity between pH 5.5 and pH 8.0 and notable activity at low temperatures. The KM for PEP and IC50 [glyphosate] values (the concentration of glyphosate that inhibited enzyme activity by 50%) of AroAA.sp were 78 µM and 1.5 mM, respectively. Furthermore, site-directed mutagenesis revealed that the G100A mutant had a 30-fold increase in the IC50 [glyphosate] value; while the L105P mutant showed only 20% catalytic activity compared to wild-type AroAA.sp. The specific activity of the wild-type AroAA.sp, the G100A mutant and the L105P mutant were 7.78 U/mg, 7.26 U/mg and 1.76 U/mg, respectively. This is the first report showing that the G100A mutant of AroA displays considerably improved glyphosate resistance and demonstrates that Leu105 is essential for the enzyme's activity.