A novel R2R3-MYB transcription factor PbMYB1L of Pyrus bretschneideri regulates cold tolerance and anthocyanin accumulation.

KEY MESSAGE: PbMYB1L enhances the cold tolerance and anthocyanin accumulation of transgenic Arabidopsis by regulating the expression of genes related to the cold-responsive genes pathway and anthocyanin synthesis pathway. MYB transcription factors (TFs) have been demonstrated to play diverse roles in plant growth and development. In the present study, we identified a novel R2R3-MYB transcription factor, PbMYB1L, from the peel of 'Red Zaosu' pear (Pyrus bretschneideri), which was induced by cold stress and acted as a positive regulator in anthocyanin biosynthesis. Notably, the transgenic Arabidopsis lines exhibited enhanced tolerance to cold stress. Compared to the Arabidopsis wild-type plants, the transgenic lines displayed longer primary roots and reduced reactive oxygen species (ROS) levels including O2-, hydrogen peroxide (H2O2), and malondialdehyde (MDA). Furthermore, significant upregulation of key cold-responsive genes AtCBF1, AtCBF2, AtCBF3, AtCBF4, and AtKIN1 was observed in the transgenic plants under cold stress conditions compared to wild type. Arabidopsis plants overexpressing PbMYB1L had significant anthocyanin accumulation in leaves after cold treatment with quantitative results indicating higher expression of anthocyanin structural genes compared to wild type. These findings suggest that PbMYB1L not only plays a vital role in conferring cold tolerance but also acts as a crucial regulator of anthocyanin biosynthesis.

An effective method for establishing a regeneration and genetic transformation system for Actinidia arguta.

The all-red A. arguta (Actinidia arguta) is an anthocyanin-rich and excellent hardy fruit. Many studies have focused on the green-fleshed A. arguta, and fewer studies have been conducted on the all-red A. arguta. Here we reported a regeneration and Agrobacterium-mediated transformation protocol by using leaves of all-red A. arguta as explants. Aseptic seedling leaves of A. arguta were used as callus-inducing materials. MS medium supplemented with 0.3 mg·L-1 2,4-D and 1.0 mg·L-1 BA was the optimal medium for callus induction of leaves, and medium supplemented with 3 mg·L-1 tZ and 0.5 mg·L-1 IAA was optimal for adventitious shoot regeneration. The best proliferation medium for adventitious buds was MS + 1.0 mg·L-1 BA + 0.3 mg·L-1 NAA. The best rooting medium was 1/2MS + 0.7 mg·L-1 IBA with a 100% rooting rate. For the red flesh hardy kiwi variety 'Purpurna Saduwa' (A. arguta var. purpurea), leaves are receptors for Agrobacterium (EHA105)-mediated transformation. The orthogonal experiment was used for the optimization of each genetic transformation parameter and the genetic transformation of the leaves was 21% under optimal conditions. Our study provides technical parameters for applying genetic resources and molecular breeding of kiwifruit with red flesh.

Transcriptomic Analysis and the Effect of Maturity Stage on Fruit Quality Reveal the Importance of the L-Galactose Pathway in the Ascorbate Biosynthesis of Hardy Kiwifruit (Actinidia arguta).

Hardy kiwifruit (Actinidia arguta) has recently become popular in fresh markets due to its edible skin and rich nutritional value. In the present study, different harvest stages of two A. arguta cultivars, 'Issai' and 'Ananasnaya' ("Ana"), were chosen for investigating the effects of maturity on the quality of the fruit. Interestingly, Issai contained 3.34 folds higher ascorbic acid (AsA) content than Ana. The HPLC method was used to determine the AsA content of the two varieties and revealed that Issai had the higher content of AsA and DHA. Moreover, RNA sequencing (RNAseq) of the transcriptome-based expression analysis showed that 30 differential genes for ascorbate metabolic pathways were screened in Issai compared to Ana, which had 16 genes down-regulated and 14 genes up-regulated, while compared to the up-regulation of 8 transcripts encoding the key enzymes involved in the L-galactose biosynthesis pathway. Our results suggested that AsA was synthesized mainly through the L-galactose pathway in hardy kiwifruit.

Genome-Wide Investigation of G6PDH Gene in Strawberry: Evolution and Expression Analysis during Development and Stress.

As one of the key enzymes in the pentose phosphate pathway (PPP), glucose-6-phosphate dehydrogenase (G6PDH) provides NADPH and plays an important role in plant development and stress responses. However, little information was available about the G6PDH genes in strawberry (Fragaria × ananassa). The recent release of the whole-genome sequence of strawberry allowed us to perform a genome-wide investigation into the organization and expression profiling of strawberry G6PDH genes. In the present study, 19 strawberry G6PDH genes (FaG6PDHs) were identified from the strawberry genome database. They were designated as FaG6PDH1 to FaG6PDH19, respectively, according to the conserved domain of each subfamily and multiple sequence alignment with Arabidopsis. According to their structural and phylogenetic features, the 19 FaG6PDHs were further classified into five types: Cy, P1, P1.1, P2 and PO. The number and location of exons and introns are similar, suggesting that genes of the same type are very similar and are alleles. A cis-element analysis inferred that FaG6PDHs possessed at least one stress-responsive cis-acting element. Expression profiles derived from transcriptome data analysis exhibited distinct expression patterns of FaG6PDHs genes in different developmental stages. Real-time quantitative PCR was used to detect the expression level of five types FaG6PDHs genes and demonstrated that the genes were expressed and responded to multiple abiotic stress and hormonal treatments.

Comparative changes of health-promoting phytochemicals and sugar metabolism of two hardy kiwifruit (Actinidia arguta) cultivars during fruit development and maturity.

Introduction: Hardy kiwifruit (Actinidia arguta) has an extensive range of nutritional and bioactive compounds and has been valued as a great resource for kiwifruit breeding. A better understanding of the dynamic changes of the composition and accumulation of nutritional compounds during fruit development and ripening is required before genetic or cultural improvements can be targeted. Methods: In the present study, the phytochemical analysis of two A. arguta cultivars 'Yilv' and 'Lvmi-1' showed that they comprised different morphology, with a higher fruit diameter while a lower vertical fruit diameter of 'Lvmi-1' compared with 'Yilv'. The antioxidant capacity of both cultivars decreased during the maturity time and showed no significant difference between them. Furthermore, although glucose gradually increased during the maturity time, the predominant sugar composition was speculated to be fructose in 'Lvmi-1' fruit while sucrose in 'Yilv' fruit at the early fruit developmental stages. Moreover, the predominant acids in 'Yilv' and 'Lvmi-1' were citric acid followed by quinic acid, malic acid, and oxalic acid. The expression of sugar- and starch-related genes encoding the crucial enzymes suggested different changes in 'Yilv' and 'Lvmi-1'. Notably, a subsequent correlation analysis showed a significant positive correlation between sucrose phosphate synthase (SPS) expression and glucose in 'Yilv', fructokinase (FK) expression, and starch content in 'Lvmi-1', implying their vital roles in sugar and starch accumulation. By contrast, a significant negative correlation between FK expression and fructose in 'Lvmi-1' fruit was observed. Results and Discussion: In summary, our results provide supplementary information for the dynamic changes of nutritional compounds and antioxidant capacity during hardy kiwifruit maturity time and give a clue for exploring the mechanism of sugar and starch accumulation in hardy kiwifruit.