Plant-based FRET biosensor discriminates environmental zinc levels.

Heavy metal accumulation in the environment poses great risks to flora and fauna. However, monitoring sites prone to accumulation poses scale and economic challenges. In this study, we present and test a method for monitoring these sites using fluorescent resonance energy transfer (FRET) change in response to zinc (Zn) accumulation in plants as a proxy for environmental health. We modified a plant Zn transport protein by adding flanking fluorescent proteins (FPs) and deploying the construct into two different species. In Arabidopsis thaliana, FRET was monitored by a confocal microscope and had a 1.4-fold increase in intensity as the metal concentration increased. This led to a 16.7% overall error-rate when discriminating between a control (1µm Zn) and high (10mm Zn) treatment after 96h. The second host plant (Populus tremula×Populu salba) also had greater FRET values (1.3-fold increase) when exposed to the higher concentration of Zn, while overall error-rates were greater at 22.4%. These results indicate that as plants accumulate Zn, protein conformational changes occur in response to Zn causing differing interaction between FPs. This results in greater FRET values when exposed to greater amounts of Zn and monitored with appropriate light sources and filters. We also demonstrate how this construct can be moved into different host plants effectively including one tree species. This chimeric protein potentially offers a method for monitoring large areas of land for Zn accumulation, is transferable among species, and could be modified to monitor other specific heavy metals that pose environmental risks.

Color Shade Nets Affect Plant Growth and Seasonal Leaf Quality of Camellia sinensis Grown in Mississippi, the United States.

Shading modifies the microenvironment and can provide plants with some protection from frequent heat, drought, frost, and hail induced by climate change and has the potential to improve plant growth, yield, and quality. Tea (Camellia sinensis) is an ancient plant originating from tropical and subtropical regions and prefers to grow in partial shade under the forest canopy. The emerging tea industry in the United States (US) requires research support on establishing tea fields in novel environmental conditions as well as on producing high-quality tea products. This study investigated the effects of black, blue, and red shade nets on tea plant growth and seasonal leaf qualities in the southeastern US with a humid subtropical climate. When compared to no-shade control, black, blue, and red shade nets increased plant growth index (PGI), net photosynthetic rate (P n), and stomatal conductance (g s), decreased air and leaf surface temperatures in summer, and reduced cold damage in winter. No significant difference was found among the black, blue, and red shade nets on tea plant growth. Varying contents of total polyphenols, carbohydrates, free amino acids, L-theanine, gallic acid, caffeine, and catechins in fresh tea leaves were observed among different shade treatments and harvesting seasons. 69.58% of the variations were depicted in a biplot by principal component analysis. Red shade was considered helpful for improving green tea quality by increasing the content of L-theanine and free amino acids in tea leaves collected in spring and fall when compared to no-shade control.

Poplar maintains zinc homeostasis with heavy metal genes HMA4 and PCS1.

Perennial woody species, such as poplar (Populus spp.) must acquire necessary heavy metals like zinc (Zn) while avoiding potential toxicity. Poplar contains genes with sequence homology to genes HMA4 and PCS1 from other species which are involved in heavy metal regulation. While basic genomic conservation exists, poplar does not have a hyperaccumulating phenotype. Poplar has a common indicator phenotype in which heavy metal accumulation is proportional to environmental concentrations but excesses are prevented. Phenotype is partly affected by regulation of HMA4 and PCS1 transcriptional abundance. Wild-type poplar down-regulates several transcripts in its Zn-interacting pathway at high Zn levels. Also, overexpressed PtHMA4 and PtPCS1 genes result in varying Zn phenotypes in poplar; specifically, there is a doubling of Zn accumulation in leaf tissues in an overexpressed PtPCS1 line. The genomic complement and regulation of poplar highlighted in this study supports a role of HMA4 and PCS1 in Zn regulation dictating its phenotype. These genes can be altered in poplar to change its interaction with Zn. However, other poplar genes in the surrounding pathway may maintain the phenotype by inhibiting drastic changes in heavy metal accumulation with a single gene transformation.

Screening Tea Cultivars for Novel Climates: Plant Growth and Leaf Quality of Camellia sinensis Cultivars Grown in Mississippi, United States.

The United States (U.S.) consumed over 80 billion servings of tea, approximately 3.8 billion gallons, in the year of 2018. With the vast majority of tea demand being met by importation, the United States became the third largest tea importer worldwide after Russia and Pakistan. As demand for domestically produced tea increases and growers expressing increasing interest in growing and producing tea, tea production became an emerging industry in the United States. Compared to major tea producing countries with centuries of growing history, tea production in the United States is limited and requires research support in many aspect of tea production including selecting suitable cultivars adapted to local climatic conditions. This study evaluated nine tea cultivars, including 'BL1,' 'BL2,' 'Black Sea,' 'Christine's Choice,' 'Dave's Fave,' 'Large Leaf,' 'Small Leaf,' 'Sochi,' and 'var. assamica,' for plant growth, leaf morphological characteristics, cold tolerance, and leaf biochemical compositions when grown in Mississippi United States with a subtropical climate. The nine tested cultivars had varying plant growth indices (PGI) and varying degrees of cold tolerance to freezing temperatures in winter, but resumed healthy growth the following spring. 'BL2' showed the highest PGI of 104.53 cm by February 2019, which might be helpful toward suppressing weed and early establishment of tea plantation. The nine cultivars also showed varying leaf characteristics in terms of leaf length, width, area, fresh and dry weights, and new shoot weight. There existed a diversity in leaf biochemical composition including soluble solids, carbohydrates, total polyphenols (TP), free amino acids (AA), L-theanine and caffeine among the nine cultivars and among different harvesting seasons of spring, summer, and fall within a certain cultivar. The nine cultivars in this study generally grow well in local environment. All tea samples collected from nine cultivars and three seasons were considered suitable for green tea processing with low TP/AA ratios ranging from 1.72 to 3.71 in this study.

Effects of induction treatments on flowering in Populus deltoides.

Stimulation of early flowering is required to shorten breeding cycles of eastern cottonwood (Populus deltoides Bartr. ex Marsh. var. deltoides), a commercially important and fast-growing hardwood species. A series of experiments was conducted to evaluate the influence of various treatments on flowering in rooted cuttings from mature and juvenile trees. A combined treatment of water stress, root pruning and paclobutrazol was applied to 3-month-old rooted cuttings from mature trees. These cuttings had been subjected to root restriction and long days. All treated plants flowered, whereas no untreated plants formed flower buds. One-year-old rooted cuttings from juvenile trees did not flower when treated with either paclobutrazol, paclobutrazol plus water stress, paclobutrazol plus root pruning, or paclobutrazol plus girdling. This was true both under continuous or periodic growth. Assessment of the lack of flowering in juvenile trees may require an integrated approach that investigates environmental or physiological stimuli, assimilate shift, gibberellic acid type and concentration, and flowering-time gene activity in the new shoots of mature and juvenile cottonwood trees.

Shoot morphogenesis associated with flowering in Populus deltoides (Salicaceae).

Temporal and spatial formation and differentiation of axillary buds in developing shoots of mature eastern cottonwood (Populus deltoides) were investigated. Shoots sequentially initiate early vegetative, floral, and late vegetative buds. Associated with these buds is the formation of three distinct leaf types. In May of the first growing season, the first type begins forming in terminal buds and overwinters as relatively developed foliar structures. These leaves bear early vegetative buds in their axils. The second type forms late in the first growing season in terminal buds. These leaves form floral buds in their axils the second growing season. The floral bud meristems initiate scale leaves in April and begin forming floral meristems in the axils of the bracts in May. The floral meristems subsequently form floral organs by the end of the second growing season. The floral buds overwinter with floral organs, and anthesis occurs in the third growing season. The third type of leaf forms and develops entirely outside the terminal buds in the second growing season. These leaves bear the late vegetative buds in their axils. On the basis of these and other supporting data, we hypothesize a 3-yr flowering cycle as opposed to the traditional 2-yr cycle in eastern cottonwood.