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plants Article LED Lights Promote Growth and Flavonoid Accumulation of Anoectochilus roxburghii and Are Linked to the Enhanced Expression of Several Related Genes Do Thi Gam 1,2, Phan Hong Khoi 2, Pham Bich Ngoc 1,3 , Ly Khanh Linh 1, Nguyen Khac Hung 1, Phan Thi Lan Anh 2, Nguyen Thi Thu 2, Nguyen Thi Thu Hien 1, Tran Dang Khanh 4,5 and Chu Hoang Ha 1,3,* 1 2 3 4 5 Received: 8 September 2020; Accepted: 6 October 2020; Published: 12 October 2020 Institute of Biotechnology, Vietnam Academy of Science and Technology (VAST), 18 Hoang Quoc Viet, Nghia Do, Cau Giay, Hanoi 100000, Vietnam; honggam@htd.vast.vn (D.T.G.); pbngoc@ibt.ac.vn (P.B.N.); lykali139@gmail.com (L.K.L.); hungkhacibt@gmail.com (N.K.H.); nguyenhien2111@gmail.com (N.T.T.H.) Center for High Technology Development, Vietnam Academy of Science and Technology (VAST), 18 Hoang Quoc Viet, Nghia Do, Cau Giay, Hanoi 100000, Vietnam; phkhoi@htd.vast.vn (P.H.K.); lananh52cnsh@gmail.com (P.T.L.A.); nminhthu02@gmail.com (N.T.T.) Vietnam Academy of Science and Technology (VAST), Graduate University of Science and Technology, 18 Hoang Quoc Viet, Nghia Do, Cau Giay, Hanoi 100000, Vietnam Agricultural Genetics Institute, Pham Van Dong, North Tu Liem, Hanoi 123000, Vietnam; tdkhanh@vaas.vn Center for Agricultural Innovation, Vietnam National University of Agriculture, Hanoi 131000, Vietnam * Correspondence: chuhoangha@ibt.ac.vn; Tel.: 84-9121-75636 Abstract: Anoectochilus roxburghii is a wild edible species and has been traditionally used for a wide range of diseases in many countries. Our research aims to find the optimal light-emitting diode (LED) lighting conditions to improve the growth and development of A. roxburghii seedling at the acclimation stage. Two-month-old explants were cultured under the various lighting conditions including red (R), blue (B), BR (one blue: four red), BRW151 (one blue: five red: one white), BRW142 (one blue: four red: two white), and fluorescent lamp (FL). The results showed that the lighting conditions not only affect the growth and morphology of plants but also the accumulation of total flavonoids. Single wavelengths (B or R LED) inhibited the growth and secondary biosynthesis of A. roxburghii, while the BR LED showed an enhancement in both growth and biomass accumulation. A. roxburghii plants were grown under BR LED light has average plant height (7.18 cm), stem diameter (17.6mm), number of leaves (5.78 leaves/tree), leaf area (4.67 cm2), fresh weight (0.459 g/tree), dry matter percentages (11.69%), and total flavonoid (1.811 mg/g FW) is considered to be superior to FL lamps and other LEDs in the experiment. This indicates that both blue and red wavelengths are required for the normal growth of A. roxburghii. To learn more about how light affects flavonoid biosynthesis, we evaluated the expression of genes involved in this process (pal, chs, chi, and fls) and found that BR LED light enhances the expression level of chi and fls genes compared to fluorescent lamps (1.18 and 1.21 times, respectively), leading to an increase in the flavonoid content of plant. Therefore, applying BR LED during in vitro propagation of A. roxburghii could be a feasible way to improve the medicinal value of this plant. Keywords: Anoectochillus roxburghii; chalcone synthase; flavonol synthase; flavonoid; LED light 1. Introduction Anoectochilus roxburghii (Orchidaceae) is one of the most valuable medicinal plants and is mainly distributed in Asian countries including Vietnam, India, Japan, Taiwan, and China. A. roxburghii Plants 2020, 9, 1344; doi:10.3390/plants9101344 www.mdpi.com/journal/plantsPDF Image | LED Lights Promote Growth and Flavonoid Accumulation of Anoectochilus roxburghii
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