scholarly journals Improvement of growth and bacoside production in Bacopa monnieri through induced autotetraploidy with colchicine

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e7966 ◽  
Author(s):  
Phithak Inthima ◽  
Kawee Sujipuli

Bacopa monnieri is a medicinal herb that is increasing in demand in Thailand. However, the lack of high-bacoside cultivars has limited pharmaceutical utilization and production. Here, chromosome doubling in B. monnieri was attempt to improve biomass and bacoside content in its seedling. Nodal segments were treated with colchicine (0, 0.025, 0.05, 0.075, 0.1, and 0.5% w/v) for 24 or 48 h before transferring to multiple shoot induction medium (1/2 MS medium supplemented with 0.2 mg L−1 BAP). Of 326 tested clones, 18 and 84 were mixoploids and autotetraploids, respectively. The highest autotetraploid-induction percentage (14.6%) was found after treated with 0.5% (w/v) colchicine, and 48 hours exposure. From 28 selected autotetraploid clones, 21 and 13 have significantly higher fresh and dry weight compared to the diploid clone, respectively. The maximum fresh and dry weight of autotetraploid plants was 2.8 and 2.0-time higher than diploid plants, respectively. Moreover, the maximum total bacoside content (1.55 mg plant−1) was obtained from an autotetraploid plant, which was 2.3-fold higher than the level in diploid plants. These novel autotetraploids have the potential to be developed as resources for value-added improvements in the medicinal and pharmaceutical industries.

2018 ◽  
Vol 44 (3) ◽  
pp. 459-463
Author(s):  
PK Roy

An efficient protocol was developed for in vitro mass propagation of Paulownia tomentosa (Thunb.) Steud. using shoot tip and leaf segment explants from field grown plant. Different concentrations and combinations of BAP, Kn, zeatin and NAA were used for multiple shoot regeneration. Among two types of explants, leaf segment produced the highest number of shoots per explant (12 ± 0.4) when they were cultured on MS supplemented with 3.0 mg/l Kn and 0.5 mg/l NAA. Addition of 10% CW to above mentioned medium increased the number of shoots (18) per culture. Shoot tip explants also produced multiple shoots in the same medium, but their performance was not good as leaf segment explants. For shoot elongation, 100 mg/l urea was more effective when added with best shoot induction medium. Shoots rooted well in halfstrength MS supplemented with 2.0 mg/l NAA, within 12 - 15 days. Regenerated plantlets were successfully acclimatized and established in poly bag containing a mixture of soil and compost in 2:1 ratio. About 90% plantlets survived under open field conditions.


Agrociencia ◽  
2020 ◽  
Vol 54 (3) ◽  
pp. 387-399
Author(s):  
Soledad Mora-Vásquez ◽  
Silverio García-Lara ◽  
Edgardo J. Escalante-Vázquez ◽  
Guy A. Cardineau

Plant tissue culture provides an alternative approach to improve the quality of soybean (Glycine max (L.) Merrill) cultivars. This study was undertaken to analyze the susceptibility of Mexican soybean for direct shoot regeneration and to determine the critical factors that affect in vitro performance. Our hypothesis was that Mexican soybean is suitable for in vitro regeneration using a cotyledonary node as explant. The effects of the seed disinfection procedure, soaking pretreatment before germination, soybean variety, as well as the culture medium composition of the shoot induction medium, were evaluated by two split-plot statistical designs. According to the statistical analysis, the seed disinfection procedure, the soaking pretreatment before germination, and the soybean genotype were the factors that brought about a significant effect (p£0.01), while the hormones composition of the shoot induction medium did not have a significant effect. The best response for multiple shoot formation was observed using a chlorine gas seed disinfection method, 3% hydrogen peroxide soaking pretreatment, and Huasteca-100, Nainari and Suaqui soybean genotypes. A robust protocol was developed, and under these selected conditions, it is possible to obtain more than 10 shoots per explant. Well-developed plantlets were obtained after 60 d of in vitro culture.


Author(s):  
Rupesh S. Badere ◽  
Pallavi K. Rinkey

The shoot-tip explant harvested from ethyl methane sulphonate (EMS) and gamma ray (GR) mutagenized seedling was cultured over MS medium fortified with NAA and BAP for five generations to amplify the mutated sector. Mutagens reduced the regeneration efficiency of the explant and affected its plant growth regulator-dependence for multiple shoot induction. While the 12d-old shoot-tip from GR-treated seedling induced shoots with 0.5µM NAA+6.6µM BAP; that from EMS-treated seedling induced shoots with 8.8µM BAP. The present study establishes that the mutagens affect the regeneration process in the explant.


HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 461d-461
Author(s):  
Richard L. Bell ◽  
Ralph Scorza ◽  
Chinnathambi Srinivasan

An efficient regeneration/transformation system was developed for `Beurre Bosc' pear. Young leaves were harvested from in vitro shoots proliferated on a medium containing MS basal salts and 5 BAP, 0.5 μM IBA, and 0.6M3. Shoot regeneration was optimized using a modification of the medium of Chevreau and Leblay (1993). Explants were cultured on shoot induction medium contained 10 μM TDZ and 1 μM IBA for 4 weeks in the dark, and then transfered to a similar, but auxinless, regeneration medium until shoots developed, usually after an additional 4 to 8 weeks. Leaf tissues were transformed by co-cultivation for 3 days with Agrobacterium tumefaciens EHA101 carrying a pGA482 plasmid containing NPTII, GUS, and rolC genes, followed by cultivation on SIM containing 300 mg/L timentin. Putative transgenic plants were selected on shoot induction medium containing 80mg/L kanamycin, and multiplied on shoot proliferation medium. Four clones were confirmed as transgenic using the GUS histochemical assay and Southern blots for the NPTII and rolC genes. Plants of each clone have been rooted and successfully transfered to the greenhouse for further analysis of gene expression.


2019 ◽  
Vol 6 (1) ◽  
Author(s):  
Hui-Hui Su ◽  
Fei Peng ◽  
Pei Xu ◽  
Xiao-Ling Wu ◽  
Min-Hua Zong ◽  
...  

Abstract Background Glucaric acid, one of the aldaric acids, has been declared a “top value-added chemical from biomass”, and is especially important in the food and pharmaceutical industries. Biocatalytic production of glucaric acid from glucuronic acid is more environmentally friendly, efficient and economical than chemical synthesis. Uronate dehydrogenases (UDHs) are the key enzymes for the preparation of glucaric acid in this way, but the poor thermostability and low activity of UDH limit its industrial application. Therefore, improving the thermostability and activity of UDH, for example by semi-rational design, is a major research goal. Results In the present work, three UDHs were obtained from different Agrobacterium tumefaciens strains. The three UDHs have an approximate molecular weight of 32 kDa and all contain typically conserved UDH motifs. All three UDHs showed optimal activity within a pH range of 6.0–8.5 and at a temperature of 30 °C, but the UDH from A. tumefaciens (At) LBA4404 had a better catalytic efficiency than the other two UDHs (800 vs 600 and 530 s−1 mM−1). To further boost the catalytic performance of the UDH from AtLBA4404, site-directed mutagenesis based on semi-rational design was carried out. An A39P/H99Y/H234K triple mutant showed a 400-fold improvement in half-life at 59 °C, a 5 °C improvement in $$ {\text{T}}_{ 5 0}^{ 1 0} $$ T 50 10 value and a 2.5-fold improvement in specific activity at 30 °C compared to wild-type UDH. Conclusions In this study, we successfully obtained a triple mutant (A39P/H99Y/H234K) with simultaneously enhanced activity and thermostability, which provides a novel alternative for the industrial production of glucaric acid from glucuronic acid.


2010 ◽  
Vol 20 (1) ◽  
pp. 73-79 ◽  
Author(s):  
M. F. Hasan ◽  
B. Sikdar

An efficient protocol for plant regeneration through multiple shoots induction from shoot tips of Polygonum hydropiper (L.) was established. The highest percentage (96.6) of multiple shoot induction and number of shoots (9.0) per culture were found on MS supplemented with 2.0 mg/l Kn. The induced shoots were excised and inoculated on to MS contains different concentrations of IBA or NAA for rooting. The highest percentage (90.0) of root induction and the highest number of roots per shoot (12.0) was found on MS having 1.0 mg/l IBA. Well rooted plantlets were acclimated properly and transplanted in the soil under natural condition, where cent per cent plantlets survived and grew successfully. Key words:  Polygonum hydropiper, Shoot tips, In vitro propagation D.O.I. 10.3329/ptcb.v20i1.5970 Plant Tissue Cult. & Biotech. 20(1): 73-79, 2010 (June)


2017 ◽  
Vol 27 (4) ◽  
pp. 530-538 ◽  
Author(s):  
Sarah J. Pethybridge ◽  
Niloofar Vaghefi ◽  
Julie R. Kikkert

Table beet (Beta vulgaris ssp. vulgaris) production in New York is increasing for direct sale, use in value-added products, or processing. One of the most important diseases affecting table beet is cercospora leaf spot (CLS) caused by the fungus Cercospora beticola. CLS causes lesions on leaves that coalesce and leads to premature defoliation. The presence of CLS may cause buyer rejection at fresh markets. Defoliation from CLS may also result in crop loss because of the inability to harvest with top-pulling machinery. The susceptibility of popular table beet cultivars (Boldor, Detroit, Falcon, Merlin, Rhonda, Ruby Queen, and Touchstone Gold) to CLS was tested using C. beticola isolates representative of the New York population. Two trials were conducted by inoculating 6-week-old plants in the misting chamber. A small-plot replicated field trial was also conducted to examine horticultural characteristics of the cultivars. In the misting chamber trials, disease progress measured by the area under the disease progress stairs (AUDPS) was not significantly different between the red cultivars, Detroit and Ruby Queen, and was significantly higher in ‘Boldor’ than the other yellow cultivar Touchstone Gold. In the field trial, the number of CLS lesions per leaf at the final disease assessment and AUDPS were significantly lower in cultivar Ruby Queen than others and not significantly different between the yellow cultivars. The dry weight of roots was not significantly different among cultivars at first harvest (77 days after planting). At 112 days after planting, the dry weight of roots was significantly higher in cultivar Detroit than Rhonda and Boldor. Leaf blade length and the length:width ratio were cultivar-dependent, which may facilitate selection for specific fresh markets. Significant associations between canopy reflectance in the near infrared (IR) (830 nm), dry weight of foliage, and number of CLS lesions per leaf suggested that this technique may have utility for remote assessment of these variables in table beet research. Implications of these findings for the management of CLS in table beet are discussed.


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