The phytohormone abscisic acid (ABA) plays a crucial role not only in fruit development and ripening, but also in adaptive responses to biotic and abiotic stresses. In these processes, the actions of ABA are under the control of complex regulatory mechanisms involving ABA metabolism, signal transduction, and transport. The endogenous ABA content is determined by the dynamic balance between biosynthesis and catabolism, processes which are regulated by 9-cis-epoxycarotenoid dioxygenase (NCED) and ABA 8'-hydroxylase (CYP707A), respectively. ABA conjugation by cytosolic UDP-glucosyltransferases, or release by β-glucosidases, is also important for maintaining ABA homeostasis. Recently, multiple putative ABA receptors localized at different subcellular sites have been reported. Among these is a major breakthrough in the field of ABA signalling-the identification of a signalling cascade involving the PYR/PYL/RCAR protein family, the type 2C protein phosphatases (PP2Cs), and subfamily 2 of the SNF1-related kinases (SnRK2s). With regard to transport, two ATP-binding cassette (ABC) proteins and two ABA transporters in the nitrate transporter 1/peptide transporter (NRT1/PTR) family have been identified. In this review, we summarize recent research progress on the role of ABA in fruit ripening, stress response, and transcriptional regulation, and also the functional verification of both ABA-responsive and ripening-related genes. In addition, we suggest possible commercial applications of genetic manipulation of ABA signalling to improve fruit quality and yields.

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The role of abscisic acid in fruit ripening and responses to abiotic stress

Phytohormone abscisic acid (ABA) induces anthocyanin biosynthesis; however, the underlying molecular mechanism is less known. In this study, we found that the apple MYB transcription factor MdMYB1 activated anthocyanin biosynthesis in response to ABA. Using a yeast screening technique, we isolated MdbZIP44, an ABA-induced bZIP transcription factor in apple, as a co-partner with MdMYB1. MdbZIP44 promoted anthocyanin accumulation in response to ABA by enhancing the binding of MdMYB1 to the promoters of downstream target genes. Furthermore, we identified MdBT2, a BTB protein, as an MdbZIP44-interacting protein. A series of molecular, biochemical, and genetic analysis suggested that MdBT2 degraded MdbZIP44 protein through the Ubiquitin-26S proteasome system, thus inhibiting MdbZIP44-modulated anthocyanin biosynthesis. Taken together, we reveal a novel working mechanism of MdbZIP44-mediated anthocyanin biosynthesis in response to ABA.

Apple bZIP transcription factor MdbZIP44 regulates abscisic acid-promoted anthocyanin accumulation.

Phytohormones and polyamines regulate plant stress responses by altering GABA pathway.

The microRNA319 (miR319) family is conserved among diverse plant species. In rice (Oryza sativa L.), the miR319 gene family is comprised of two members, Osa-miR319a and Osa-miR319b. We found that overexpressing Osa-miR319b in rice resulted in wider leaf blades and delayed development. Here, we focused on the biological function and potential molecular mechanism of the Osa-miR319b gene in response to cold stress in rice. The expression of Osa-miR319b was down-regulated by cold stress, and the overexpression of Osa-miR319b led to an enhanced tolerance to cold stress, as evidenced by higher survival rates and proline content. Also, the expression of a handful of cold stress responsive genes, such as DREB1A/B/C, DREB2A, TPP1/2, was increased in Osa-miR319b transgenic lines. Furthermore, we demonstrated the nuclear localization of the transcription factors, OsPCF6 and OsTCP21, which may be Osa-miR319b-targeted genes. We also showed that OsPCF6 and OsTCP21 expression was largely induced by cold stress, and the degree of induction was obviously repressed in plants overexpressing Osa-miR319b. As expected, the down-regulation of OsPCF6 and OsTCP21 resulted in enhanced tolerance to cold stress, partially by modifying active oxygen scavenging. Taken together, our findings suggest that Osa-miR319b plays an important role in plant response to cold stress, maybe by targeting OsPCF6 and OsTCP21.

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MicroRNA319 Positively Regulates Cold Tolerance by Targeting OsPCF6 and OsTCP21 in Rice (Oryza sativa L.)

This review contains functional roles of MYB transcription factors in the transcriptional regulation of anthocyanin biosynthesis in horticultural plants This review describes potential uses of MYB TFs as tools for metabolic engineering for anthocyanin production Anthocyanins (ranging from red to blue) are controlled by specific branches of the anthocyanin biosynthetic pathway and are mostly visible in ornamentals, fruits, and vegetables In the present review, we describe which R2R3-MYB transcription factors (TFs) control the transcriptional regulation of anthocyanin structural genes involved in the specific branches of the anthocyanin biosynthetic pathway in various horticultural plants (eg, ornamentals, fruits, and vegetables) In addition, some MYBs responsible for anthocyanin accumulation in specific tissues are described Moreover, we highlight the phylogenetic relationships of the MYBs that suppress or promote anthocyanin synthesis in horticultural crops Enhancement of anthocyanin synthesis via metabolic genetic engineering of anthocyanin MYBs, which is described in the review, is indicative of the potential use of the mentioned anthocyanin-related MYBs as tools for anthocyanin production Therefore, the MYBs would be suitable for metabolic genetic engineering for improvement of flower colors, fruit quality, and vegetable nutrients

Roles of R2R3-MYB transcription factors in transcriptional regulation of anthocyanin biosynthesis in horticultural plants

Leaf-chewing insects are important pests that cause yield loss and reduce seed quality in soybeans (Glycine max). Breeding soybean varieties that are resistant to leaf-chewing insects can minimize the need for insecticide use and reduce yield loss. The marker gene for QTL-M, Glyma.07g110300 (LOC100775351) that encodes a UDP-glycosyltransferase (UGT) is the major determinant of resistance against leaf-chewing insects in soybean; it exhibits a loss of function in insect-resistant soybean germplasms. In this study, Agrobacterium-mediated transformation introduced the CRISPR/Cas9 expression vector into the soybean cultivar Tianlong No. 1 to generate Glyma.07g110300-gene mutants. We obtained two novel types of mutations, a 33-bp deletion and a single-bp insertion in the GmUGT coding region, which resulted in an enhanced resistance to Helicoverpa armigera and Spodoptera litura. Additionally, overexpressing GmUGT produced soybean varieties that were more sensitive to H. armigera and S. litura. Both mutant and overexpressing lines exhibited no obvious phenotypic changes. The difference in metabolites and gene expression suggested that GmUGT is involved in imparting resistance to leaf-chewing insects by altering the flavonoid content and expression patterns of genes related to flavonoid biosynthesis and defense. Furthermore, ectopic expression of the GmUGT gene in the ugt72b1 mutant of Arabidopsis substantially rescued the phenotype of H. armigera resistance in the atugt72b1 mutant. Our study presents a strategy for increasing resistance against leaf-chewing insects in soybean through CRISPR/Cas9-mediated targeted mutagenesis of the UGT genes.

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CRISPR/Cas9-Mediated Targeted Mutagenesis of GmUGT Enhanced Soybean Resistance Against Leaf-Chewing Insects Through Flavonoids Biosynthesis

A LEAFY/FLORICAULA (LFY/FLO) homolog PpLFL (PrunuspersicaLEAFY/FLORICAULALike) gene was isolated from axillary buds of peach (Prunus persica (L.) Batsch. cv. Bayuecui) during flower induction period. The open reading frame of PpLFL spanned 1,248 bp, encoding a putative protein of 415 amino acid residues, which was with high similarity (50.48 %–84.69 %) to other FLO/LFY inferred proteins from different species. The spatial expression patterns of PpLFL were detected in axillary buds during the periods of flower induction by using immunohistolocalisation. The results showed that PpLFL gene was mainly expressed during flower induction time, and also in leaf and petal promordia at the SAM. For further functional analysis, the PpLFL was constitutively expressed in the Arabidopsis lfy mutant background, and the results showed that overexpression of PpLFL under the control of CaMV 35S promoter can accelerate flowering and give rise to normal flower organs. Our results suggest that PpLFL might play an important role in flower induction, and could act as a functional flower meristem identity gene in peach.

Identification and Characterization of PpLFL, a Homolog of FLORICAULA/LEAFY in Peach ( Prunus persica )

Rapeseed (Brassica napus L.) is an important oil crop species and plays a crucial role in supplying edible oil worldwide. However, rapeseed production in the field is often severely inhibited due to nitrogen (N) deficiency. Metabolites play key roles in plant growth and resistance to environmental stress, but little is known about the differential synthesis and accumulation of metabolites underlying rapeseed adaptation to N deficiency. Here, we studied the phenotypic response and used LC–electrospray ionization (ESI), ESI–MS/MS, and widely untargeted metabolomic approaches to detect differences in rapeseed under normal N (HN) and N-deficient (LN) conditions. The results showed that N deficiency severely inhibited rapeseed shoot growth and promoted rapeseed root architectural changes under LN conditions. In total, 574 metabolites were detected, and there were 175 and 166 differentially accumulated metabolites in the leaves and roots between the HN and LN conditions, respectively. The significantly differentially accumulated metabolites were involved in four primary metabolic pathways, namely, sucrose, phenylalanine, amino acid, and tricarboxylic acid cycle metabolism. Notably, we found that plant hormones have distinct accumulation patterns in rapeseed and coordinate to play crucial roles in both maintaining growth and protecting against damage from plant disease under HN and LN conditions. Moreover, our results indicated that flavonoid compounds, especially anthocyanins and rutin, may play important roles in increasing root cell resistance to oxidative damage and soil pathogen infections. Overall, this work provides valuable information for understanding the overall metabolite changes in rapeseed under N deficiency conditions, which may be beneficial for improving and producing new varieties of rapeseed capable of high yields under low N conditions.

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Metabolic Profiles Reveal Changes in the Leaves and Roots of Rapeseed (Brassica napus L.) Seedlings under Nitrogen Deficiency

The role of polyol transporters in stress tolerance in plants have been elucidated by many studies. Sorbitol transporter genes MdSOT3, MdSOT4 and MdSOT5 in apple plants, which are important for sorbitol loading and unloading, are regulated by drought stress. To further confirm the role of sorbitol transporters in stress tolerance, the constructs harboring MdSOT3 and MdSOT5 genes were introduced into wild type Arabidopsis plants (Col-0) and the Arabidopsis transformed with MdSOT3 or MdSOT5 performed higher drought stress tolerance compared to WT. In order to further understand how sorbitol transporters are involved in drought tolerance in apple plants, upstream regions of sorbitol transporter genes were isolated from apple plant source leaves by Anchored PCR from genomic DNA obtained, and then were used to drive expression of the GUS reporter in tobacco plants. The results showed that the longest fragments of MdSOT3 and MdSOT5 promoters induced the highest GUS activity under drought stress conditions. Additionally, fragments of these promoters that contain cis-acting elements known to be involved in stress response also induced GUS activity under drought stress. Taken together, our data suggest that increased MdSOT3 and MdSOT5 activity, through cis-acting elements in the promoters of these genes, play important roles in imparting tolerance to drought in micropropagated apple plants.

Isolation and characterization of two sorbitol transporter gene promoters in micropropagated apple plants (Malus × domestica) regulated by drought stress

Xinjie Shen

Papers

CRISPR/Cas9-Mediated Targeted Mutagenesis of GmUGT Enhanced Soybean Resistance Against Leaf-Chewing Insects Through Flavonoids Biosynthesis

Identification and Characterization of PpLFL, a Homolog of FLORICAULA/LEAFY in Peach ( Prunus persica )

Metabolic Profiles Reveal Changes in the Leaves and Roots of Rapeseed (Brassica napus L.) Seedlings under Nitrogen Deficiency

Isolation and characterization of two sorbitol transporter gene promoters in micropropagated apple plants (Malus × domestica) regulated by drought stress

Malus sieversii (Ledeb.) Roem-derived plant growth-related protein, and coding gene and application thereof