Effects of <i>Bacillus</i> <i>amyloliquefaciens</i> QST713 on Photosynthesis and Antioxidant Characteristics of Alfalfa (<i>Medicago sativa</i> L.) under Drought Stress

Drought stress is a prevalent abiotic stress that adversely affects multiple physiological processes in plants, especially their photosynthetic capacity. Application of plant growth–promoting rhizobacteria (PGPR) has been considered as an eco-friendly strategy to ameliorate the deleterious effects of drought stress on plants. The present study was carried out to investigate the effects of Bacillus amyloliquefaciens QST713 on plant growth, leaf relative water content (RWC), photosynthesis processes, photosynthetic pigment content and antioxidant enzyme activities in two alfalfa varieties, Galalxie Max (drought-tolerant) and Saidi 7 (drought-sensitive) under drought conditions. The results showed that drought stress significantly declined plant biomass production, RWC, photosynthetic pigment content (Chl a, Chl b and carotenoids) and photosynthetic gas exchange parameters (transpiration rate (Tr), stomatal conductance (Gs) and intercellular CO2 concentration (Ci)), whereas it enhanced the enzymatic activity of peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) in both cultivars. In contrast, the inoculation of the bacillus strain QST713 was more effective on plant growth, showing higher plant biomass production compared to the non-inoculated plants under drought stress. Moreover, the application of QST713 significantly promoted the content of RWC, the accumulation of chlorophyll content and the activities of antioxidant enzymes as well as enhanced the photosynthetic capacity of alfalfa seedlings under drought stress. These results suggest that QST713 could be considered as a promising bio-inoculant for plants exposed to environmental stresses

Two B-Box Proteins, MaBBX20 and MaBBX51, Coordinate Light-Induced Anthocyanin Biosynthesis in Grape Hyacinth

Floral colour is an important agronomic trait that influences the commercial value of ornamental plants. Anthocyanins are a class of flavonoids and confer diverse colours, and elucidating the molecular mechanisms that regulate their pigmentation could facilitate artificial manipulation of flower colour in ornamental plants. Here, we investigated the regulatory mechanism of light-induced anthocyanin biosynthesis during flower colouration in grape hyacinth (Muscari spp.). We studied the function of two B-box proteins, MaBBX20 and MaBBX51. The qPCR revealed that MaBBX20 and MaBBX51 were associated with light-induced anthocyanin biosynthesis. Both MaBBX20 and MaBBX51 are transcript factors and are specifically localised in the nucleus. Besides, overexpression of MaBBX20 in tobacco slightly increased the anthocyanin content of the petals, but reduced in MaBBX51 overexpression lines. The yeast one-hybrid assays indicated that MaBBX20 and MaBBX51 did not directly bind to the MaMybA or MaDFR promoters, but MaHY5 did. The BiFC assay revealed that MaBBX20 and MaBBX51 physically interact with MaHY5. A dual luciferase assay further confirmed that the MaBBX20–MaHY5 complex can strongly activate the MaMybA and MaDFR transcription in tobacco. Moreover, MaBBX51 hampered MaBBX20–MaHY5 complex formation and repressed MaMybA and MaDFR transcription by physically interacting with MaHY5 and MaBBX20. Overall, the results suggest that MaBBX20 positively regulates light-induced anthocyanin biosynthesis in grape hyacinth, whereas MaBBX51 is a negative regulator

Identification and Functional Analysis of a Flavonol Synthase Gene from Grape Hyacinth

Flavonols are important copigments that affect flower petal coloration. Flavonol synthase (FLS) catalyzes the conversion of dihydroflavonols to flavonols. In this study, we identified a FLS gene, MaFLS, expressed in petals of the ornamental monocot Muscari aucheri (grape hyacinth) and analyzed its spatial and temporal expression patterns. qRT-PCR analysis showed that MaFLS was predominantly expressed in the early stages of flower development. We next analyzed the in planta functions of MaFLS. Heterologous expression of MaFLS in Nicotiana tabacum (tobacco) resulted in a reduction in pigmentation in the petals, substantially inhibiting the expression of endogenous tobacco genes involved in anthocyanin biosynthesis (i.e., NtDFR, NtANS, and NtAN2) and upregulating the expression of NtFLS. The total anthocyanin content in the petals of the transformed tobacco plants was dramatically reduced, whereas the total flavonol content was increased. Our study suggests that MaFLS plays a key role in flavonol biosynthesis and flower coloration in grape hyacinth. Moreover, MaFLS may represent a new potential gene for molecular breeding of flower color modification and provide a basis for analyzing the effects of copigmentation on flower coloration in grape hyacinth