PDF Publication Title:
Text from PDF Page: 011
Plants 2020, 9, 1344 11 of 15 (Ushiyama Denki, LTD). Cross and longitude sections were directly observed under DN-200 Digital Microscope. Cells width and length were measured by the Scope Image 9.0 software (by Bioimager, Inc.). Values shown in Table 1 are the mean of 03 replicates; each replication contains the samples from 5 plants in each treatment with 5 sections/node/plant. 4.4. Determination of Total Flavonoids Contents The total flavonoid content was determined by the colorimetric method of Marinova et al., 2005, and Sultana et al. [63,64]. The principle of the method is based on the coloration of flavonoid compounds with AlCl3 in an alkaline medium. One gram of fresh leaf sample was crushed homogeneously in liquid nitrogen, then extracted with absolute ethanol at the ratio of 1:3. The sample was further extracted on an ultrasonic machine for 30 min, then centrifuged to recover the ethanol solution. The extracted ethanol obtained from 3 repeats were combined and adjusted to equal volumes (10 mL) with ethanol. We shook well the mixture of 1 mL of the obtained extract, 4 mL of water and 0.3 mL of 5% NaNO2, left the mixture at room temperature for 5 min, then added 0.3 mL of 10% AlCl3. After 6 min, we added 2 mL of 1 M NaOH solution, mixed well, and adjusted the volume with water to 10 mL. The absorbance of the mixtures was recorded at 510 nm by UV/VIS Spectrophotometer Camspec M108. The total flavonoid contents were calculated based on the quercetin calibration curve, performed under the same conditions. The results were expressed in mg/g of fresh leaf [65]. 4.5. Expression Analysis of Several Flavonoid Biosynthesis-Related Genes Three samples of each light condition were used for total RNA extraction. The third leaves of 2-month-old plants were collected at the same time, after 6 h exposed to the light regime in a daily 12 h light cycle. Total RNA was isolated from A. roxburghii leaves by using TRizolTM Reagent (InvitrogenTM) following manual instruction and then transcribed to cDNAs using RevertAid First Strand cDNA Synthesis Kit (ThermoFisher Scientific). The expressional levels of flavonoid biosynthesis-related genes (pal, chs, chi, and fls) together with actin2 as housekeeping gene was determined by real-time PCR system (Rotor-Gene Q, QUIAGEN). PCR was conducted by using SYBR Green (SsoAvanced Universal SYBR Green Supermix, BioRad) by following manufacturing instruction. PCR thermal program was performed as following: pre-denature 95 ◦C/3 min; 40 cycles of denature 95 ◦C/10 s; annealing 57 ◦C/20 s; extension 72 ◦C/20 s. DNA melting curve was determined by ramp heating from 65 ◦C to 95 ◦C and holding within 5 s at 95 ◦C for the analysis. The different relative level of gene expression between lighting conditions was determined based on the 2(-Delta Delta C(T)) method [66]. The experiment is considered to be different when processing ANOVA statistics of 03 replications with p-value < 0.05. Specific primers for analysis were described by Yang et al. [67]; Zhang et al. [68], and Zhang et al. [69,70]. The specific gene primers involved in flavonoids biosynthesis-related genes expression are presented in Supplementary Table S1. 4.6. Statistical Analysis StartGraphic XV (Statpoint Technologies, Inc., Warrenton, VA, USA) was used for all statistical analyses. Data were analyzed using one-way ANOVA analysis. Differences between the means of each treatment were detected by using Duncan’s multiple range test with p < 0.05. 5. Conclusions In summary, light source influenced not only plant growth and morphogenesis but also metabolites accumulation. This paper showed that A. roxburghii required both blue and red wavelengths for normal development. Plants grew under BR light (1 blue diodes with 4 red diodes) obtained the highest stem diameter, leaf area, and biomass. Especially, BR light could promote the accumulation of flavonoid properly due to enhancing expression of several critical genes involved in the flavonoid biosynthesis pathway. This paper provides an effective method that uses LED light to improve the development and flavonoid accumulation in A. roxburghii.PDF Image | LED Lights Promote Growth and Flavonoid Accumulation of Anoectochilus roxburghii
PDF Search Title:
LED Lights Promote Growth and Flavonoid Accumulation of Anoectochilus roxburghiiOriginal File Name Searched:
plants-09-01344.pdfDIY PDF Search: Google It | Yahoo | Bing
Cruise Ship Reviews | Luxury Resort | Jet | Yacht | and Travel Tech More Info
Cruising Review Topics and Articles More Info
Software based on Filemaker for the travel industry More Info
The Burgenstock Resort: Reviews on CruisingReview website... More Info
Resort Reviews: World Class resorts... More Info
The Riffelalp Resort: Reviews on CruisingReview website... More Info
CONTACT TEL: 608-238-6001 Email: greg@cruisingreview.com (Standard Web Page)