6.

CONCLUSION AND FUTURE PERSPECTIVES

The results of the present study describe the role of biochemical

and molecular markers in the identification of the best cultivar tenera variety of

oil palm Elaeis guineensis. The data confirms the application of the molecular

method, RAPD-SCAR, in identifying the variety tenera at vegetative phase

and at the seedling stage. This goal has been achieved by two phases of

investigation in the broad sense. The first is the search for a biochemical

marker by focusing the enzyme-mediated pathway of shell synthesis, the

morphological marker used for identification of tenera variety. This approach

is quiet different from the earlier reports on oil palm shell synthesis, since the

production and utilization of r!02 in lignin synthesis has been discussed in

connection with shell formation in oil palm for the first time. The other is to find

out a consistent molecular marker related to shell synthesis.

The correlation of 02- and H202 in shell formation was

demonstrated by assaying the marker enzymes NADPH Oxidase, super oxide

dismutase and peroxidase and the preferential role of these enzymes, during

lignin synthesis in dura and tenera, in controlling the shell thickness was

established figure 31 ). The quantification data of 02- ions and l-b02 further

110
 Cell System O2

02-

l~

[POX I ICADI
-4------- Monolignols..-- Cinnamaldehydes

Fig. 31. H20 2 mediated pathway of shell

formation in oil palm fruit
SOD - Super oxide dismutase, POX - Peroxidase,
CAD - Cinnamyl alcohol {NADPH} dehydrogenase
supported the assay data. The varied level of K102 in the endocarp region of

dura and tenera provided a clear indication of their difference in the shell

thickness. Being a toxic component, the scavenging of H202 from the cell

system directly or indirectly has become an inevitable metabolic need for the

survival of the plant. The association of 1102 in the formation and deposition

of lignin in the shell points to two aspects: the scavenging of toxicity by the

removal of 1102 and to the formation of endocarp itself. This in turn suggests

that H202 has a decisive role in the thickness of the shell. The higher amount

of H202 and the thick shell in dura compared to that of tenera very well

supports this hypothesis.

Irrespective of the difference in activity of NADPH oxidase,

super oxide dismutase, peroxidase and cinnamyl alcohol (NADPH)

dehydrogenase between mesocarp and endocarp of both dura and tenera,

the activity as a whole in the fruits did not infer satisfactory difference between

dura and tenera. Hence these enzymes could not be treated as biochemical

markers for differentiating the two varieties. So the study of detecting genetic

polymorphism of dura and tenera by RAPD and subsequent advancement of

developing a SCAR primer has elevated the whole investigation to a higher

level for solving a vexing problem of oil palm breeders and researchers. The

application of SCAR primer in detecting tenera was confirmed by repeated

trials in mature palms as well as seedlings. Considering the positive effects of

SCAR primer for detecting the tenera variety at academic level, more trials

111
are warranted for ascertaining the practical viability of the study, so that it will

be helpful to oil palm breeders and farmers. With the present status, the

investigation can be extended at molecular level by collecting germ plasm of

tenera from different plantations, using variable parameters viz age of palm,

water, and temperature so that the purity of the primer can be checked. More

over, an RNA finger printing can be done more extensively between the

varieties dura and pisifera by OORT- PCR method in order to determine the

differential expression of genes during shell synthesis.

Any sort of molecular attempt to control the activity of enzymes

related to shell synthesis, by regulating the specific genes, has very much

relevance in the production of a thin shelled oil palm. But this vision of

hypothesis has its own impediments, while considering the functional roles of

these enzymes as well as the physiological roles of their end products. For

instance, the peroxidase enzyme has multifaceted functions in the plant

metabolic system other than lignin formation and the knockdown of peroxidase

may produce other physiological problems. Even though cinnamyl alcohol

(NADPH) dehydrogenase (CAD) enzyme has a pertinent role in the formation

of shell, the manipulation of this enzyme gene would not be a positive thinking

unless the production of ROS has been regUlated. This leaves super oxide

dismutase (SOD) as an ideal target enzyme for controlling the production of

H202 by either manipulating the SOD gene or interfering the promoter, which

would lead to the formation of an oil palm fruit with a thin shell.

112