Development of duck weed transformation technique for biological application

Name: Aornpilin Jaiprasert

LCSH: Burapha University.Faculty of Science

LCSH: Duckweeds

LCSH: Duckweeds -- Utilization

LCSH: Genetic transformation -- Technique

LCSH: Transgenic plants

Abstract: Plants are important as global energy and food sources, especially during the situation where the world population is rapidly increasing. Though the genetically modified crops such as maize and soybean are commercial available, these crops encounter several limitations, including long harvesting period, taking up land space, and requiring an extensive investment. Consequently, constructing genetically modified plants that can be cultivated in limited space and short-time life cycles is desirable. Duckweeds are one of the promising choices due to their rapid biomass duplication and high carbohydrate and protein contents. Nevertheless, the basic information and gene transformation technique in duckweeds are poorly described. Therefore, the main objectives of this research are studying the fundamental of duckweed biology, analyzing their biochemical composition and developing the simple method for genetic transformation of duckweeds. Three species of duckweeds found in Burapha University, Chon-Buri, Thailand, were characterized, including Spirodela polyrhiza, Lemna aequinoctialis and Wolffia globosa. They were subsequently surfaced sterilized using NaClO and successfully cultured axenically in the laboratory. When grown in Hoagland’s E medium, their doubling times were 2.4 days (L. aequinoctialis), 3.2 days (S. polyrhiza), and 3.6 days (W. globosa), respectively. Interestingly, of all duckweeds, S. polyrhiza was able to accumulate carbohydrate and protein content upto 40.7% and 31.4%, respectively. In contrast, the highest level of carbohydrate content in L. aequinoctialis was only 6.9% compensated for its fastest growth. Furthermore, addition of salicylic acid, a plant growth regulator, to the culture media triggered flowering in S. polyrhiza. The floral structure of S. polyrhiza was incomplete, lacking of petal and sepal, but was a perfect flower v having one pistil and two stamens. Agrobacterium-mediated transformation was performed to genetically modify S. polyrhiza by agroinfiltration of its turions, specialized fronds. Intriguingly, vacuum infiltration for 5 min yielded the highest transformation frequency as shown by the detection frequency of Bar and Egfp up to 75% and the observation of GFP fluorescence signals in the first generation of transgenic duckweeds (T1). Nevertheless, the transgenes were apparently lost during vegetative proliferation as shown by the reduction of the detection frequency of Bar and Egfp to 50% and 0%, respectively, in T2 (the second generation of transgenic duckweeds). Altogether, the striking superiority of S. polyrhiza, including rapid growth, high carbohydrate and protein content, inducible flowering, a simple transformation protocol, and an availability of its genomic DNA sequence, will make this duckweed as a versatile tool in biotechnology to cope with the world’s challenging problems, especially food and energy security, in the near future.

Burapha University. Library

Address: CHONBURI

Email: buulibrary@buu.ac.th

Name: Salil Chanroj

Role: Principal advisor

Created: 2018

Modified: 2020-03-05

Issued: 2020-03-05

วิทยานิพนธ์/Thesis

application/pdf

CallNumber: Th 584.64 Ao638D

eng

DegreeName: Master of Science

Level: Master's Degree

Descipline: Biological Science

Grantor: Burapha University

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