Production Of Planting Stock From Kacip Fatimah [Labisia Pumila (Bi.) F.Vill & Neves] Through Cuttings

Labisia pumila (Bl.) F.Vill & Naves (Myrsinaceae) has been traditionally used by Malay women in parturition. Three different varieties of this species were identified and found in Malaysia: viz pumila, alata and lanceolata. However, two of its varieties i.e pumila and alata have been commonly used in traditional medicine and researched as they are more readily available than lanceolata. The increase in logging activities and encroachment in forests caused a decline in the production of L. pumila from its natural stands. This problem is further aggravated by its slow growth in natural forests. Moreover, it is difficult to obtain planting material from the wild. In order to address the demand of L. pumila, it is necessary to find alternative methods of production, specifically vegetative propagation by cuttings, to ensure regular supply of its planting stock. This study aims to explore the possibility of mass producing L. pumila using different propagation systems (misting and non misting), plant parts (stem, petiole and leaf) of two varieties (alata and pumila) and rooting media (river sand, sawdust and 1:1 v/v mixture of river sand and sawdust). This study utilized a Strip-Split Plot (SSPD) in a Randomised Complete Block Design (RCBD). This experiment was conducted in FRIM, Kepong’s, nursery. Results after three months propagation indicated that there were significant differences in all treatments with regards to root and shoot growth. Cuttings of var. alata produced better than var. pumila in terms of root and shoot growth. Propagation in misting systems produced cuttings with higher rooting ability (84%) compared to the ones in non-misting system (72%). However, cuttings in the non-misting system showed higher shooting ability (28%). Cuttings grown in 100% river sand produced higher root and shoot abilities (87% and 26%) than those raised on mixture of 1:1 river sand and sawdust (76% and 24%) and sawdust alone (72% and 23%). Propagation using stem cuttings were found to be the best compared to leaf and petiole cuttings. Based on the elongation of root cells, stem cuttings of variety alata was found to be better than pumila. The elongation of root cell cuttings in those three media, showed a young root primordium elongated through the cortex from parenchyma phloem cell. These root cells were bigger size grown on sawdust and mixture of river sand and sawdust compared to river sand alone. Again, stem cuttings showed the best growth performance than the other plant parts based on height and diameter. In addition, upon chemical analysis of these cuttings, the flavonoids and resorcinol-derivatives content increased in mother plant of var. alata. However, stem rooted cuttings of var. alata had decreased in resorcinol-derivatives, which means it will give less adverse effects to humans compared to var. pumila cuttings. The results of this research demonstrated that L. pumila var. alata stem cuttings could be possibly propagated vegetatively through rooting of cuttings with favorable treatments such those raised on river sand medium under the misting propagation system

Bio-organic fungicide of Catharanthus roseus stems extract inhibit the growth of Fusarium oxysporumon Capsicum annum seedling

Fusarium oxysporum is a well-known fungus that causes a major commercial plant disease in the world. Due to the issue, the antifungal activity of Catharanthus roseus stems extract as bio-organic fungicide against F. oxysporumon Capsicum annum seedling was studied. The spore suspensions of F.oxysporum and C. roseusstems extract were prepared to study the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) tests in the laboratory. The application of C. roseus stems extract at the concentrations of 100, 500, 1,000, 1,500 and 2,000 µg/mL against F. oxysporum were included in the in-vitro study. For in-vivo test, F. oxysporum on C. annum seedlings were applied with the plant extract at concentrations of 1,000, 1,500 and 2,000 µg/mL in greenhouse study. The result showed that the extractwith2,000 µg/mL has higher significant difference (p<0.05) inhibited the growth of F. oxysporum plant fungal compared to other concentrations of MIC tested. The MFC test indicated that day nine was proven to have high negative impact of the fungal than day six. In in-vivo study, the antifungal activity showed 100% effect of disease injury for the growth of C. annum plant species in green house. The result also showed that the concentration of C. roseus stems extract at 2,000 µg/mL has significantly higher (p<0.05) activity against F. oxysporum on the seedlings compared to other concentrations. Thus, the study indicated that C. roseus stems extract has novelty of bio-organic compounds that contribute to the development of new antifungal agents to protect crop plants from fungal disease which also safe to environmental ecology compared to other commercial chemical fungicide which is highly used nowadays

Effect of a misting system and rooting media of Labisia pumila cuttings

Labisia pumila is one of the most popular and potent ingredients used in Malay traditional herbal. However, logging activities and forest encroachment reduced the productivity of L. pumila in natural stands. In addition, the species is slow growth and its scarcity of wildings in natural forests. Due to the problems, a study on L. pumila cuttings to examine the possibility of raising planting stocks was studied. The experiment was done at FRIM's nursery with a Split-Split Plot (SSPD) in a Randomized Complete Block Design (RCBD) involving two propagation systems (misting and nonmisting), three rooting media (river sand, sawdust and 1:1 mixture of river sand and sawdust) and three plant parts (stem, petiole and leaf). Results indicated significant differences (p ≤ 0.05) in all treatments with regards to their root and shoot development. Cuttings propagated in misting system produced better rooting ability (84%) than the non-mist system (72%). However, cuttings propagated in non-mist system showed better shoot growth (29%) than the ones raised in the misting system (20%). Cuttings grown in river sand produced higher root and shoot abilities (87% and 26%) than those raised on river sand and sawdust mix (1:1) (76% and 24%) and in sawdust (72% and 23%). Stem cuttings performed best compared to leaf and petiole cuttings. In conclusion, L. pumila var. alata stem cuttings could be possibly propagated vegetatively through rooting of cuttings with favorable treatments such those raised on river sand medium under the misting propagation system

Isaria fumosorosea and Metarhizium anisopliae for controlling Atteva sciodoxa (Lepidoptera: Yponomeutidae), a pest of Eurycoma longifolia

Tiger moth, Atteva sciodoxa (Lepidoptera: Yponomeutidae), is a major pest of tongkat ali, Eurycoma longifolia (Simaroubaceae). To find a safe and effective method for controlling the pest, two indigenous entomopathogenic fungi, Isaria fumosorosea and Metarhizium anisopliae, isolated from bagworms Pteroma pendula (Lepidoptera: Psychidae), were bioassayed against the pest. The larvae were separately sprayed with concentrations of 1 × 102 to 1 × 105 conidia mL-1 of each fungal isolate. Both fungi were pathogenic to third instar larvae of A. sciodoxa. However, M. anisopliae was more virulent than I. fumosorosea. The median effective concentrations for M. anisopliae and I. fumosorosea were 4.23 × 10 3 and 8.24 × 104 conidia mL-1 respectively. The median infective times ranged from 4.3 to 10.3 days for M. anisopliae and 7.6 to 16.3 days for I. fumosorosea. Metarhizium anisopliae killed 48 to 88% larvae while I. fumosorosea, 26 to 62% larvae for the lowest and highest concentrations respectively, 10 days after treatment. Spraying of M. anisopliae at 2 × 107 conidial mL-1 suspension reduced the population of A. sciodoxa attacking 2-year-old saplings up to 89%, 7 days after treatment. This study indicates the potential of M. anisopliae for controlling A. sciodoxa

Catharanthus roseus extract as bio-fungicide for controlling Fusarium oxysporum on selected vegetable seedlings

Catharanthus roseus extracts were studied to evaluate their potentials replacing chemical fungicide which give a bad impact on human health and cause environmental pollution problems. The samples of C. roseus were extracted with different extraction solvents including dichloromethane (DCM), acetone, ethanol, and methanol at 5, 10, 15 and 20 mg/mL concentrations. The solvents without the plant extract were used as the controls. After 6 days of incubation, the inhibition zone of the fungal pathogen on PDA media was measured. The extracts were significantly effective (p ≤ 0.05) in limiting the antifungal activities. The DCM extract of C. roseus was the most effective against Fusarium oxysporum with 8.06 mm compared to acetone (0.055 mm), ethanol (0.15 mm), methanol (0.41 mm), and water (0.06 mm). Among of the concentrations, 20 mg/mL gave the best effect to control the fungal pathogen compared to 5, 10 and 15 mg/mL. The C. roseus extract was also effective in controlling F. oxysporum on the selected vegetable seedlings based on leaf number and disease suppression (%) results. However, without treated with the extract; mortality due to F. oxysporum increased. The C. roseus extract was effective and may to be developed as a bio-fungicide agent to control F. oxysporum in the field

Diversity of endophytic fungi isolated from different plant parts of Acacia mangium, and antagonistic activity against Ceratocystis fimbriata, a causal agent of Ceratocystis wilt disease of A. mangium in Malaysia

Acacia mangium is an important wood for commercial products especially pulp and medium-density fibreboard. However, it is susceptible to Ceratocystis fimbriata infection, leading to Ceratocystis wilt. Therefore, the present work aimed to (i) establish the diversity of endophytic fungi in different plant parts of A. mangium,and (ii) evaluate the antifungal potentials of the isolated and identified endophytic fungi against C. fimbriata. Endophytic fungal identification was conducted by PCR amplification and sequencing of the internal transcribed spacer 1 (ITS1) and ITS4 regions of nuclear ribosomal DNA. A total of 66 endophytic fungi were successfully isolated from different parts of A. mangium; leaf (21), stem (13), petiole (12), root (9), flower (6), and fruit (5). The endophytic fungal isolates belonged to Ascomycota (95.5%) and Zygomycota (4.5%). For Ascomycota 13 genera were identified: Trichoderma (28.6%), Nigrospora (28.6%), Pestalotiopsis (12.7%), Lasiodiplodia (9.5%), Aspergillus (6.3%), Sordariomycetes (3%), and Neopestalotiopsis, Pseudopestalotiopsis, Eutiarosporella, Curvularia, Fusarium, Penicillium, and Hypoxylon each with a single isolate. For Zygomycota, only Blakeslea sp. (5%) was isolated. Against C. fimbriata, Trichoderma koningiopsis (AC 1S) from stem, Nigrospora oryzae (AC 7L) from leaf, Nigrospora sphaerica (AC 3F) from the flower, Lasiodiplodia sp. (AC 2 U) from fruit, Nigrospora sphaerica (AC 4P) from petiole, and Trichoderma sp. (AC 9R) from root exhibited strong inhibition for C. fimbriata between 58.33 to 69.23%. Thus, it can be concluded that certain endophytic fungi of A. mangium have the potential to be harnessed as anti-Ceratocystis agent in future biotechnological applications

Screening of antifungal compound isolated from catharanthus roseus L. (pink) for biological control of selected plant diseases

Diseases of rubber (Hevea brasiliensis) caused by pathogens, Rigidoporus microporus, Ganoderma philippii and Phellinus noxius. Fusarium oxysporum,F. solani and Colletotrichum gloeosporioides on chilli (Capsicum annuum) are currently being kept under control with chemical fungicides. However, these fungicides have been shown to have hazardous effects to humans and the ecosystems. To address these problems, the search for an effective and environmentally safe compound to control these harmful pathogens is highly warranted. Thus, in this study, antifungal compounds from selected plants, Aglaia argentea, A. leucophylla, A. grandis, A. odoratissima, A. variisquama,Cassia alata, Catharanthus roseus, Derris elliptica and Tinospora baenzigeri were screened for their effectiveness in controlling the growth of selected fungal pathogens on seedlings. The result showed that C. roseus extract was the most effective in inhibiting the growth of pathogens as the extract contains various antifungal compounds such as phenolics, alkaloids, essential oils and flavonoids. Although the extract of C. roseus contains abundant bioactive compounds, detailed studies on their biological activities on those fungal pathogens have yet to be reported. The C. roseus extracted with dichloromethane (DCM) showed the greatest inhibition zone diameter in controlling R. microporus and F. oxysporum at the values of 11.29 mm and 8.10 mm, respectively, compared to other selected medicinal plant extracts. The C. roseus extract assessed based on conidia and hyphae growth of F. oxysporum and R. microporus, respectively showed minimum inhibition concentration (MIC) and minimum fungicidal concentration (MFC). MIC and MFC results showed that the extract at 270 μg/mL with inhibition value of 0.0 μg/mL was the most effective in controlling the growth of F. oxysporum and R. microporus compared to 200, 140, 70 and 10 μg/mL. The antifungal compound isolated from C. roseus extract was determined through thin layer chromatography (TLC), high performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR) analysis. Each C. roseus of DCM extracts was marked as CRD5a, CRD5b, CRD5c, CRD5d, CRD5e, CRD5f and CRD5g. The TLC results showed that all of the C. roseus extracts peak with red in colour at Rf= 0.61 at 366 nm wavelength, except for CRD5g. The CRD5d extract was the most effective against G. philippii and R. microporus with inhibition zones of 3.5 and 1.9 mm, respectively, compared to other extracts. However, CRD5g extract was the most effective against F. oxysporum with a value of 3.0 mm compared to other extracts. HPLC results also showed the major peak is at 210 nm. The CRD5d extract isolated contained single compound such as ursolic acid after being detected by NMR analysis. The compound was effective to control R. microporus and G. philippii with inhibition zone values of 4.0 and 3.0 mm, respectively. In addition, the efficacy of C.roseus extracts against F. oxysporum and R. microporus was assessed based on healthy effects percentage of the chilli and rubber seedlings, respectively,by assessing the symptoms on leaves and roots. The assessment was based on disease incident (DI%) and disease suppression (DS%). The DI% results showed that an extract at 2,000 μg/mL was the most effective in controlling F.oxysporum on chilli seedlings with value of 0.0%, compared to 1000 and 1500 μg/mL with values of 60 and 80%, respectively. The DS% results also showed that the extract was most effective at 2000 μg/mL in controlling the growth of R.microporus on rubber seedlings with a value of 100% compared to 1500 and 1000 μg/mL with a value of 0.0%. In conclusion, C. roseus extracted with DCM contains an effective toxin that is detrimental on the plant pathogenic fungi. The C. roseus of DCM extract isolated contains ursolic acid and the compound effective against R. microporus and G. philippii. Hence, C. roseus extract should also be developed as a biofungicide for controlling R. microporus and F.oxysporum on rubber and chilli, respectively

In vivo evaluation of Catharanthus roseus L. (pink) extracts against Rigidoporus microporus on rubber (Hevea brasiliensis) seedlings

Hevea brasiliensis (rubber) is an important species in rubber industries. The species is often attacked by Rigidoporus microporus in the plantations and chemical fungicides have been widely used to control this fungus. However, fungicides contribute to human health problems and environmental pollutions. To address the issues, bio-fungicide from the plant sources is an innovative alternative to replace the chemical fungicide. Thus, the objectives of the study were to investigate the effect of Catharanthus roseus extract in different concentrations against R. microporus on rubber seedlings. The efficacy of C. roseus extracts against R. microporus was assessed based on healthy effects percentage of the rubber seedlings, by assessing the symptoms on leaves and roots. The assessment was based on disease index (DI) and disease suppression (DS%). The DI results showed, at 2000 μg/mL was the most effective against R. microporus on rubber seedlings with level disease index of level 0 (0%) compared to 1500 and 1000 μg/mL with level 2 (26 - 50%), and level 4 (76 - 100%), respectively. The DS% results also showed that the extract was most effective at 2000 μg/mL in controlling the growth of R. microporus on rubber seedlings with a value of 100% compared to 1500 and 1000 μg/mL with a value of 0.0%. Hence, C. roseus extract might be suitable use as a biofungicide for controlling R. microporus on rubber crops

Bioantifungal activity of selected medicinal plants extracts against roots rot of fungal disease

Root rot disease of fungal such as Ganoderma philippii, Phellinus noxius and Rigidoporus microporus are well known destructive fungus to plant root. The in vitro antifungal screening of selected medicinal plants was studied against the disease. The twelve of Malaysian medicinal plants including the leaves of Aglaia argentea, A. leucophylla, A. grandis, A. odorata, A. odoratissima, A. varrisquama, Alium sativum (bulbs) and Cassia alata, Catharanthus roseus stems and leaves, Derris elliptica leaves and Tinospora baenzigeri stems were extracted using different types of solvents extraction i.e, dichloromethane (DCM), acetone and methanol at the concentration of 20 mg/mL. The extracts were studied for antifungal activities against three species of fungal disease including G. Philippii, P. noxius and R. microporus. The antifungal activities of the extracts were determined by the presence or absence of fungal inhibition zone growth on Potato dextrose agar (PDA). The extracts shows a significant results but varying in their antifungal activities on the selected fungal. The DCM and acetone extracts of C. roseus stems had the highest antifungal activities against R. microporus fungus compared to methanolic extract. On the other hand, acetone extracts of A. argentea leaves also gave the highest antifungal activities against G. philippii compared to other extracts. However, all of the extracts didn’t show any inhibition zone on P. noxius culture. In general, the DCM extracts of C. roseus stems contain the most of bio-antifungal of active compounds against R. microporus of fungal disease