The somatic embryogenesis rate was very high, proving that the embryogenesis ability of Dendrobium orchids is very sensitive to the culture medium supplemented with high BA concentration and the ability to generate somatic embryos was even higher when we added a combination of BA and NAA to the culture medium, showing the following results:
- In V 2 MS + 7mg/l BA environment, number of embryos: 7.67
- In MS + 7mg/l + 0.5mg/l NAA medium , number of embryos: 7.78
- In the y 2 MS + 10mg/l BA environment , the number of embryos: 8.67
- In medium y 2 MS + 10mg/l BA + 0.5mg/l NAA, number of embryos: 9.11
in vitro
Thus, BA and NAA are suitable for somatic embryogenesis in in vitro culture.
*number of protocorms:
Based on table 4.5, it shows that there are differences between environmental factors on the ability to
protocorm formation ability at 60 days of culture.
In the control medium (1/2 MS), the number of protocorms formed was 1.44, the ability to form protocorms increased in mediums supplemented with high BA, because BA has the role of stimulating cell division, so the higher the concentration of BA, the higher the ability to divide cells. Therefore, the number of protocorms formed is high. However, the role of BA is strongly stimulated when we combine BA with NAA, so the medium supplemented with BA and NAA often shows high ability to form protocorms.
This shows that BA and NAA play a role in the ability to create protocorm after 60 days of culture.
*quantity of shoots:
Based on table 4.5, there is a difference between the culture media on the ability to form buds. At this stage, the ability to form buds of Dendrobium is also not high, because the culture sample mainly generates somatic embryos and creates protocorms.
The number of shoots formed was high in culture media without somatic embryogenesis and protocorm formation such as: l Á MS + lmg/1 BA the number of shoots was 5.11, Vi MS + 3mg/l BA the number of shoots was 6.11 and in the medium Vi MS + lmg/1 BA + 0.5mg/l NAA the number of shoots was
6.11. In other experimental environments, the number of shoots decreased.
Thus, at the 60-day culture stage, BA and NAA did not have a significant effect on the ability to form buds.
*Bud height:
Shoot height always decreased in media supplemented with high BA or BA combined with NAA, because BA only stimulates cell division but not growth.
Table 4.9: Dendrobium orchid shoots after 60 days of in vitro culture on medium supplemented with BA .
Figure 4.10: Dendrobium orchid shoots after 60 days of in vitro culture on medium supplemented with BA and NAA.
After 90 days of culture, the number of shoots increased significantly, the number of protocorms and
Somatic embryos all develop into buds thus growth and development of buds.
In this stage we only observe the birth
Table 4.6: Effects of BA and NAA on the growth and development of Dendrobium orchid shoots in vitro after 90 days of culture
Solution | number | number of leaves | SORE / | Height | ||
ViMS MS + lmg/1 BA MS + 3mg/l BA MS + 5mg/l BA MS + 7mg/l BA MS + 10mg/l BA Vi MS+1mgABA+0.5mgANAA Vi MS+3mg/lBA+0.5mg/lNAA Vi MS+5mgABA+0.5mgflNAA Vi MS+7mg/lBA+0.5mg/INAAAA Vi 1.64 MS+10mg/lBA+ | 4.11 A 5.22*® 6.44 BC 7.22 CD 8.00 DE 9.33 F 6.56 c 7.22 CD 8.00 DE 9.11® | 0.40^ | 2.00 ^ 2.15® 2.12® 2.13® 1.68 A 1.67 A 2.11® 2.33® 2.29® 2.06® | 1.89 F 0.77 D 0.66 CD 0.39 A 0.34 A 0.30 A 1.23 E 1.86° 0.64 BCD 0.49 ABC 0.98 | A | 2.83 F 2.51F 1.73™ 1.52 ®c 1.30 AB 1.21 “ 2.12® 1.97 DE 1.91DE 1.30^ |
cv% | 0.88% | 1% | 1.84% | 1.1% | ||
The difference is statistically significant at the 0.05 level. | ||||||
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Comment:
*number of cards:
The classification test table shows that there are differences between environmental factors on the shoot formation process in the 90-day culture period.
The number of shoots increased very high on the medium supplemented with high BA concentration and increased even higher when we added a combination of BA and NAA to the culture medium:
- In MS Vi medium + 7mg/l BA number of shoots: 8.00
- In MS + 7mg/l BA + 0.5mg/l NAA medium , number of shoots: 9.11
- In MS + 10mg/l BA medium, number of shoots: 9.33
- In the medium of Vi MS + 10mg/1 BA + 0.5mg/l NAA, number of shoots: 10.22
Thus, after 90 days of culture, BA and NAA have a great influence on the ability to form buds.
*number of leaves, roots and shoot height:
- Number of leaves: For Dendrobium orchids , similar to Cymbidium , the higher the BA concentration, the number of leaves does not increase and may decrease.
- Number of roots: Not only in MS Vi medium , roots formed but also in other mediums, although not as high as in the control medium (1/2 MS). It shows that Dendrobium orchids are sensitive to NAA in root formation even at low concentrations of NAA.
- Shoot height: Party 4.6 showed that shoot height was highest in Vi MS medium and gradually decreased in high BA supplemented medium.
BA=5+NAA=0 3 BA=7+NAA=0 3 BA=Flood+NAA=05
Figure 4*12: Dendrupium orchid shoots after 90 days of in vitro culture on medium supplemented with BA and NAA
4.2.2. Experiment 2: Effects of TDZ and NAA on in vitro culture of Dendrobium orchid
After 60 days of culture, the experimental results showed that all treatments gave
good results
In the medium supplemented with TDZ or TDZ combined with NAA, the ability to form somatic embryos, protocorms and buds was strongly stimulated.
Table 4.7: Effects of TDZ and NAA on the ability to form somatic embryos, protocorms and buds of Dendrobium orchids after 60 days of in vitro culture
Somatic embryogenesis | Number | Number | protocorm | / Number of shoots | / sample Shoot height (cm) | ||
ViMS 0A MS+0.05mg/l TDZ 4.56 B MS+0.1mg/l TDZ 6.00 c y2 MS+0.5mg/l TDZ 7.44 D y 2 MS+lmg/1 TDZ 8.89 E MS+0.05mg/l TDZ+0.5mg/l 4.44 B NAA % MS+0.1mg/l TDZ+0.5mg/l NAA 5.00 BC % MS+0.5mg/l TDZ+0.5mg/l NAA 8.44 DE y2 MS+lmg/l TDZ+0.5mg/l NAA 9.44 E | 2.00 A 4.56 BC 5.56 BC 6.89 DE 7.33 B 4.33 CD 5.78 E 9.33 B 10.44 A | 2.89 ^ 4.56 c 2.22 A 2.56 A 3.00 ^ 3.89 BC 5.00 CD 6.11 D 2.56 A | 1.56 c 1.21 BC 0.87 ^ 0.67 A 0.59 A 1.22 BC 0.93 ^ 0.72 A 0.66 A | ||||
cv% | 1.1% | 1.33% | 1.3% | 2% | |||
*Values followed by letters in the same column with different characters indicate The difference is statistically significant at the 0.05 level. | |||||||
Comment:
Through statistical testing to classify the treatments of the above experiment, it shows that after 60 days of in vitro culture, there are the following manifestations:
*number of somatic embryos:
Based on table 4.7, it can be seen that the culture environment factors have different effects on the ability of Dendrobium orchids to produce somatic embryos.
Only in the control medium ( MS) did somatic embryogenesis not occur, while in the medium supplemented with TDZ, somatic embryogenesis was clearly evident. This shows that TDZ is very suitable for embryogenesis.
When we combined TDZ and NAA supplementation into the culture medium, it showed that the ability to generate somatic embryos was higher in the medium supplemented with only TDZ.
Thus, TDZ and NAA have great influence on the ability of somatic embryogenesis of Dendrobium orchids at 60 days of culture.
*number of protocorms:
Through the classification test table, it shows that only MS Patch medium gives us the lowest protocorm number of 2.00, while in the medium supplemented with high TDZ concentration, the number of protocorms increases. When we combine TDZ and NAA, it always gives effective cuttings:
- In MS + 0.5mg/l TDZ medium , number of protocorms: 6.89
- In MS + 0.5mg/l TDZ + 0.5mg/l NAA medium , number of protocorms: 9.33
- In MS Vi medium + lmg/1 TDZ number of protocorms: 7.33
- In Va MS medium + lmg/1 TDZ + 0.5mg/l NAA number of protocorms: 10.44
Thus, TDZ and NAA have great influence on protocorm formation of orchid species.
Dendrobium at 60 days culture stage.
*number of buds:
Based on table 4.7, it can be seen that at this stage, the ability to form buds is not high. In environments with high concentrations of TDZ or TDZ combined with NAA, the number of buds formed is low, because in these environments, protocorms are mainly created and somatic embryos are generated.
Like
so in
60-day culture period of TDZ, TDZ and NAA have
affect
not large enough to create a hut
*Bud height:
Table 4.7 shows that the higher the ability to form embryos and protocorms, the higher the tower shoot height. This proves that TDZ and NAA do not affect shoot development.
