Siti Nor Akmar Abdullah

Oil palm - achievements and potential.

Abstract Cultivation of the oil palm (Elaeis guineensis Jacq.) has expanded tremendously in recent years such that it is now second only to soybean as a major source of the world supply of oils and fats. Presently, Southeast Asia is the dominant region of production with Malaysia being the leading producer and exporter of palm oil. This paper reviews the various factors that have led to oil palm occupying its present position, including biological, technical, managerial, environmental, and socio-political aspects. Biological features recognised as critical to the high productivity of the crop are examined. These include its perennial and evergreen nature (giving a continuous year-round canopy cover intercepting a high proportion of incoming radiation), the year-round production of fruit bunches and the high partition of total assimilates into harvested product. Scientific and managerial aspects contributing to the success of the crop include the significant genetic improvements and production of high quality planting materials, the development and application of finely-tuned agronomic practices, the appropriate scale and efficient organisation of oil palm plantations and the continuous R&D and good infra-structural support provided in the main producing countries. The programmes of crop improvement through the utilisation of traditional breeding and selection methods, the development and benefits of vegetative propagation techniques using tissue culture and ongoing efforts to apply molecular and genetic engineering techniques to improve and modify oil composition, are reviewed. Finally, the nutritional qualities of palm oil as a healthy component of diet are briefly described.

SSRs for Marker-Assisted Selection for Blast Resistance in Rice (Oryza sativa L.)

Rice blast caused by the fungus Magnaporthe oryzae is one of the most devastating diseases of rice in nearly all rice growing areas of the world including Malaysia. To develop cultivars with resistance against different races of M. oryzae, availability of molecular markers along with marker-assisted selection strategies are essential. In this study, 11 polymorphic simple sequence repeat (SSR) markers with good fit of 1:2:1 ratio for single gene model in F2 population derived from the cross of Pongsu seribu 2 (Resistant) and Mahsuri (Susceptible) rice cultivars were analysed in 296 F3 families derived from individual F2 plants to investigate association with Pi gene conferring resistance to M. oryzae pathotype. Parents and progeny were grouped into two phenotypic classes based on their blast reactions. Chi-square test for the segregation of resistance and susceptibility in F3 generation fitted a ratio of approximately 3:1. Association of SSR markers with phenotypic trait in F3 families was identified by statistical analysis. Four SSR markers (RM413, RM5961, RM1233 and RM8225) were significantly associated with blast resistance to pathotype 7.2 of M. oryzae in rice (p ≤ 0.01). These four markers accounted for about 20% of total phenotypic variation. So, these markers were confirmed as suitable markers for use in marker-assisted selection and confirmation of blast resistance genes to develop rice cultivars with durable blast resistance in Malaysian rice breeding programmes.

Differential expression of oil palm pathology genes during interactions with Ganoderma boninense and Trichoderma harzianum.

The expression profiles of Δ9 stearoyl-acyl carrier protein desaturase (SAD1 and SAD2) and type 3 metallothionein (MT3-A and MT3-B) were investigated in seedlings of oil palm (Elaeis guineensis) artificially inoculated with the pathogenic fungus Ganoderma boninense and the symbiotic fungus Trichoderma harzianum. Expression of SAD1 and MT3-A in roots and SAD2 in leaves were significantly up-regulated in G. boninense inoculated seedlings at 21 d after treatment when physical symptoms had not yet appeared and thereafter decreased to basal levels when symptoms became visible. Our finding demonstrated that the SAD1 expression in leaves was significantly down-regulated to negligible levels at 42 and 63 d after treatment. The transcripts of MT3 genes were synthesized in G. boninense inoculated leaves at 42 d after treatment, and the analyses did not show detectable expression of these genes before 42 d after treatment. In T. harzianum inoculated seedlings, the expression levels of SAD1 and SAD2 increased gradually and were stronger in roots than leaves, while for MT3-A and MT3-B, the expression levels were induced in leaves at 3d after treatment and subsequently maintained at same levels until 63d after treatment. The MT3-A expression was significantly up-regulated in roots at 3d after treatment and thereafter were maintained at this level. Both SAD and MT3 expression were maintained at maximum levels or at levels higher than basal. This study demonstrates that oil palm was able to distinguish between pathogenic and symbiotic fungal interactions, thus resulting in different transcriptional activation profiles of SAD and MT3 genes. Increases in expression levels of SAD and MT3 would lead to enhanced resistance against G. boninense and down-regulation of genes confer potential for invasive growth of the pathogen. Differences in expression profiles of SAD and MT3 relate to plant resistance mechanisms while supporting growth enhancing effects of symbiotic T. harzianum.

Microsatellite-based evidences of genetic bottlenecks in the cryptic species “Andrographis paniculata Nees”: a potential anticancer agent

Andrographis paniculata (AP) is a medicinal plant species introduced into Malaysia. To address the genetic structure and evolutionary connectedness of the Malaysian AP with the Indian AP, a DNA sequence analysis was conducted based on 24 microsatellite markers. Out of the 24 primer sets, seven novel microsatellite primers were designed and amplified intra-specifically according to the available Indian AP sequences at the National Centre for Biotechnology Information (NCBI), where 17 of them were amplified using the cross-species strategy by employing the primers belonging to Acanthus ilicifolius Linn (Acanthaceae) and Lumnitzera racemosa Wild (Combretaceae). The primers were then applied on the Malaysian AP accessions. Sixteen of the new microsatellite loci were amplified successfully. Analysis of these microsatellite sequences, revealed some significant differences between the Indian and Malaysian AP accessions in terms of the size and type of the repeat motifs. These findings depicted the cryptic feature of this species. Despite identifying several heterozygous alleles no polymorphism was observed in the detected loci of the selected accessions. This situation was in concordance with the presence of “fixed heterozygosity” phenomenon in the mentioned loci. Accordingly, this was fully consistent with the occurrence of the genetic bottleneck and founder effect within Malaysian AP population. Apart from the amplification of new microsatellites in this species, our observations could be in agreement with the risk of genetic depletion and consequently extinction of this precious herb in Malaysia. This issue should be taken into consideration in the future studies.

Genetic Divergence and Heritability of 42 Coloured Upland Rice Genotypes (Oryzasativa) as Revealed by Microsatellites Marker and Agro-Morphological Traits

Coloured rice genotypes have greater nutritious value and consumer demand for these varieties is now greater than ever. The documentation of these genotypes is important for the improvement of the rice plant. In this study, 42 coloured rice genotypes were selected for determination of their genetic divergence using 25 simple sequence repeat (SSR) primers and 15 agro-morphological traits. Twenty-one out of the 25 SSR primers showed distinct, reproducible polymorphism. A dendrogram constructed using the SSR primers clustered the 42 coloured rice genotypes into 7 groups. Further, principle component analysis showed 75.28% of total variations were explained by the first—three components. All agro-morphological traits showed significant difference at the (p≤0.05) and (p≤0.01) levels. From the dendrogram constructed using the agro-morphological traits, all the genotypes were clustered into four distinct groups. Pearson’s correlation coefficient showed that among the 15 agro-morphological traits, the yield contributing factor had positive correlation with the number of tillers, number of panicles, and panicle length. The heritability of the 15 traits ranged from 17.68 to 99.69%. Yield per plant and harvest index showed the highest value for both heritability and genetic advance. The information on the molecular and agro-morphological traits can be used in rice breeding programmes to improve nutritional value and produce higher yields.

Gene expression of the oil palm transcription factor EgAP2-1 during fruit ripening and in response to ethylene and ABA treatments.

A cDNA encoding an ERE-binding protein (EgAP2-1) was isolated from the oil palm fruit mesocarp treated with ethylene using yeast one-hybrid assay. EgAP2-1 belongs to the AP2 subfamily of the APETALA2/ethylene-responsive factor (AP2/ERF) proteins and contains two highly conserved AP2/EREBP DNA-binding domains (DNA-BD). Sequence comparison of EgAP2-1 with other AP2 proteins revealed high conservation of the two AP2/EREBP domains and linker region among these proteins. Its protein was localized to the nucleus of onion epidermis cells and showed ERE-specific binding, transcriptional activation, and transactivation properties in yeast and in vitro. Its mRNA was highly expressed in oil palm mesocarp with elevated levels in ripening fruits but not in leaves and roots. EgAP2-1 was induced in mesocarp in response to ethylene and abscisic acid but not other hormonal stimuli, including methyl jasmonate and salicylic acid, and abiotic stresses including drought, cold, and high-salinity. Our results demonstrate a link between the regulation of EgAP2-1 expression and ethylene- and/or ABA-coordinated control of the fruit ripening and suggest a regulatory role for EgAP2-1 during fruit ripening and development in oil palm.

Biocontrol of Plant Parasitic Nematodes by Fungi: Efficacy and Control Strategies

Increasing knowledge and growing concern about the elevated cost of inorganic fertilizers or chemical pesticides with their vast applications on various crop plants has raised interest in the alternative method of plant disease protection caused by plant parasitic nematodes. These alternative methods are not only cost-effective but also eco-friendly to the environment and human health. Among the various rhizospheric microorganisms, opportunistic fungi like Paecilomyces lilacinus, Pochonia chlamydosporia, and arbuscular mycorrhizal (AM) fungi have the potential to reduce the severity of diseases caused by plant parasitic nematodes and also improved the plant growth and biomass production. This chapter provides an overview on the biocontrol potential of opportunistic as well as AM fungi on the growth and development of various crop plants. The details about the interactions between these fungi and plant parasitic nematodes have been discussed. An overview of the recent cost-effective technologies used for the mass propagation of these beneficial rhizospheric microorganisms is also discussed.

Highly efficient protocol for callogenesis, somagenesis and regeneration of Indica rice plants

In the present study, we have reported a simple, fast and efficient regeneration protocol using mature embryos as explants, and discovered its effective applicability to a range of Indica rice genotypes. We have considered the response of six varieties in the steps of the regeneration procedure. The results showed that calli were variably developed from the scutellar region of seeds and visible within 6-20 days. The highest and lowest calli induction frequency (70% and 51.66%) and number of induced calli from seeds (14 and 10.33) were observed in MR269 and MRQ74, respectively. The maximum and minimum number (7.66 and 4) and frequency of embryogenic calli (38.33% and 20%) were recorded in MR219 and MRQ74, respectively. However, the highest browning rate was observed in MR84 (87%) and the lowest rate in MRQ50 (46%). The majority of plants regenerated from embryogenic calli were obtained from MRQ50 (54%) and the minimum number of plants from MR84. In this study, the maximum numbers of plantlets were regenerated from the varieties with highest rate of embryogenic calli. Also, various varieties, including MRQ50, MR269, MR276 and MR219, were satisfactorily responding, while MRQ74 and MR84 weakly responded to the procedure. Such a simple, successful and generalized method possesses the potential to become an important tool for crop improvement and functional studies of genes in rice as a model monocot plant.

An Efficient Agrobacterium-Mediated Transformation of Strawberry cv. Camarosa by a Dual Plasmid System

An Agrobacterium-mediated transformation method was applied to introduce the luciferase reporter gene under the control of the CaMV35S promoter in the pGreen0049 binary vector into strawberry cv. Camarosa. The in vitro regeneration system of strawberry leaves to be used in the transformation was optimized using different TDZ concentrations in MS medium. TDZ at 16 µM showed the highest percentage (100%) of shoot formation and the highest mean number of shoots (24) produced per explant. Studies on the effects of different antibiotics, namely timentin, cefotaxime, carbenicillin and ampicillin, on shoot regeneration of strawberry leaf explants showed the best shoot regeneration in the presence of 300 mg/L timentin and 150 mg/L cefotaxime. Assessment of the different factors affecting Agrobacterium mediated-transformation of strawberry with the luciferase gene showed the highest efficiency of putative transformant production (86%) in the treatment with no preculture, bacterial OD600 of 0.6 and the addition of 150 mg/L cefotaxime in the pre-selection and selection media. The presence of the luciferase gene in the plant genome was verified by the luciferase reporter gene assay, nested PCR amplification and dot blot of genomic DNA isolated from the young leaves of each putatively transformed plantlet.

Isolation and expression analysis of transcripts encoding metallothioneins in oil palm

Two of the abundant transcripts encoding type 2 metallothionein (MT) proteins designated as MET2a and MET2b were selected in our previous study due to their high abundance (16.05 %) in the suppression subtractive hybridization library and their involvement in fruit development and maturation. The present study involves the isolation of the full-length cDNA encoding MET2a and MET2b from the ripening oil palm fruit mesocarp, examining their expression pattern compared to the other two previously reported type-3 MT members (MT3-A and MT3-B) in various oil palm organs including different vegetative and reproductive tissues. The full-length cDNA sequences of MET2a and MET2b were 571 and 553 bp and they were designated as EgMT2a and EgMT2b, respectively. The sequences of the EgMT2a and EgMT2b were then compared for sequence similarities in the database using both BLASTN and BLASTX programs. Their sequences were homologous (67–77 %) with several type-2 MTs in plants. All four MT encoding genes were differentially expressed in the ripening oil palm mesocarp tissues, but undetectable in the vegetative tissues examined. All MT genes examined were significantly up-regulated in the mature developmental stages of oil palm fruit mesocarp, except for EgMT2b which was expressed only at 17 weeks after anthesis. The type 2 MT proteins are related to a greater degree to the late fruit-ripening stage than the type 3 MT proteins consistent with their reported functions in homeostasis or detoxification. The findings in the present study contribute to better understanding the molecular mechanisms involved in fruit ripening in oil palm.