In vitro Regeneration and Microscopic Studies of Castanea sativa Mill.
Castanea sativa Mill. (European chestnut) belongs to the Fagaceae family and is known among multipurpose plants. It is a significant industrial crop that is appreciated and processed globally to produce goods in various industries including food, medicine, furniture, and timber industry. At prese...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2024
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| Subjects: | |
| Online Access: | http://ir.unimas.my/id/eprint/45979/3/Thesis%20Ms._SITI%20AISHAH%20ZAKERI.pdf |
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| Summary: | Castanea sativa Mill. (European chestnut) belongs to the Fagaceae family and is known
among multipurpose plants. It is a significant industrial crop that is appreciated and
processed globally to produce goods in various industries including food, medicine,
furniture, and timber industry. At present, it is considered a staple food throughout southern
Europe, Turkey, southwestern Asia and eastern Asia, which infers the essential for rapid
multiplication and higher yield production of the species. Yet, there is still a lack of research
on the propagation of C. sativa Mill. especially in Malaysia. Thus, the present study focuses
on investigating the optimum medium for the in vitro propagation of C. sativa to produce
plantlets on a large scale besides establishing the sterilization protocol for seed germination.
Initially, the selected seeds were surface sterilized and cultured on MS basal medium. After
2 months, the nodal segments of aseptic seedlings were sectioned into smaller pieces and
cultured on various treatments for shoot induction. Then, the cultures were maintained in the
culture room. Among the parameters recorded were the percentage of shoot production,
mean number of shoots produced, mean shoot height, mean number of leaves, and mean
leave length. Furthermore, the 6-month-old plantlets regenerated in vitro were subjected to
morphological and anatomical studies. The present study has discovered the sterilization
protocol with 80% seed germination. Besides, the stage of seedling growth and development
have been examined starting with the seed germination on the first week of culture, followed
by seedling emergence and the appearance of true leaves and hairy roots after one month of
culture. Additionally, direct in vitro propagation of C. sativa is influenced by either internal
or external factors. Among the main factors include plant growth regulators and plant
additives. Current findings clarified the treatments tested mostly promoted 100% shoot production whereby the explants could produce approximately 1-2 shoots per explant (≈1
shoot per explant) with 1.0-2.0 cm shoot height (≈2.0 cm) besides producing 1-4 leaves (2
leaves) and the leaves length within the range of 0.3-2.0 cm (1.0 cm) after 2 months of
culture. Analysis of the results disclosed the most recommended medium for regeneration of
the plant species in vitro was MS supplemented with CW + AC + 3.0mg/L BAP + 1.0mg/L
NAA whereby 100% explants producing shoot within 2 months, with an average of 2 shoots
per explant, 2.0 cm shoot height and 4 leaves with 1.0 cm length. Moreover, the
morphological study has revealed the stomata dispersed widely on the abaxial leaf with the
size of stomata opening, 12.45 μm length and 2.7 μm width. Meanwhile, the anatomical
study disclosed unique internal structures of the leaf blade and midrib including the vascular
bundles (phloem and xylem) and mesophyll cells (palisade and spongy). Hence, the present
findings provide some basic information and understanding on the propagation of plant
species in vitro. Nonetheless, further research should be performed to improve the protocol
for seed sterilization and shoot induction of C. sativa Mill. in vitro. In fact, more parameters
should be considered for data analysis as well as the other factors that could influence the
growth and performance of C. sativa Mill.
Keywords: European chestnut, micropropagation, seed germination, shoot induction, plant growth regulator. |
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