The effect of different soil nutrient and irrigation levels on Periwinkle (Chatharanthus sp.) at different altitudes: An Approach Study for Good Agricultural Parctice (GAP)

By: Ni Luh Watiniasih Putu Sudiarta Nyoman Semadi Antara

UNIVERSITAS UDAYANA KAMPUS BUKIT JIMBARAN BALI DESEMBER 2012 TABLE OF CONTENT

ABSTRACT I. INTRODUCTION 1

II. LITERATURE REVIEW

2.1. Catharanthus roseus 3

2.2. Catharanthus roseus alkaloid content and function 4

2.3. Good Agricultural Practice 5

III. MATERIAL AND METHOD 6

3.1. Location and Time of the research 6

3.2. Seedling preparation 7

3.3. Field Works: Planting and Caring 8

3.4. Plant water and nutrient treatments 9

3.4. Plant Harvest and Lab Works 11

IV. RESULT AND DISCUSSION 11

4.1. Soil Type and Mineral Content 11

4.2. The growth preference of C. roseus plant at different altitude 12

4.3. The growth rate of C. roseus plant at different water

and nutrient treatments 14

4.4. Good Agricultural Practice of Catharanthus roseus 16

REFERENCES ACKNOWLEDGMENT

i TABLE OF FIGURE

Figure: Page 1. The Madagaskar periwinkle (Catharanthus rosues (Linn.) Don)…………… 2 2. The Madagaskar periwinkle grow at Renon, Denpasar (a) and Pancasari, Tabanan (b)……………………………………………………… 7 4. Planting and caring the Madagascar periwinkle (C. roseus) before the water and nutrient treatments……………………………………. 8 3. Three months old seedlings of Madagaskar periwinkle (C. rosues)………… 8 5. The Madagaskar periwinkle before nutrient and water treatment was performed at Renon, Denpasar………………………………………… 9 6. The Madagaskar periwinkle before nutrient and water treatment was performed at Pancasari, Tabanan………………………………………. 9 7. The Madagascar periwinkle after water and nutrient treatment was performed for 4 weeks at Renon, Denpasar…………………………… 10 8. The Madagascar periwinkle after water and nutrient treatment was performed for 4 weeks at Pancasari, Tabanan………………………… 10 9. Harvesting C. roseus plants (a) and air-dried leaves (b)…………………… 11 10. The growing Madagascar periwinkle (C. roseus) in low altitude at Renon (a) and high altitude at Pancasari (b) before treated with different water and nutrient treatments ……………………………… 12 11. The C. roseus grows at Renon (a) where the individual plant was larger and taller than that at Pancasari (b)……………………………. 13 12. Average plant height, number of leaves, and number of flower of C. roseus grow in low (Renon) and high (Pancasari) altitudes………… 13 13. Average plant height, number of leaves and number of flower of C. rosues grow in low altitude (Renon) from different water and nutrient treatments. Note: C = Control; HW = High Water; LW = Low Water; HF = High Fertilizer; LF = Low Fertilizer………………….. 14 14. Average leaf area of C. rosues grow in low altitude (Renon) from different water and nutrient treatments. Note: C = Control; HW = High Water; LW = Low Water; HF = High Fertilizer; LF = Low Fertilizer…… 15 15. Average plant height, number of leaves and number of flower of C. rosues grow in high altitude (Pancasari) from different water and nutrient treatments. Note: C = Control; HW = High Water; LW = Low Water; HF = High Fertilizer; LF = Low Fertilizer………………………………. 15

ii The effect of different soil nutrient and irrigation levels on Periwinkle (Chatharanthus sp.) at different altitudes: An Approach Study for Good Agricultural Parctice (GAP)

ABSTRACT Madagascar periwinkle (Catharanthus roseus) is a shrubby perennial plant and widely spread in tropical area. They produce abundant plant bioactive chemical Terpenoid Indole Alkaloids (TIAs) and two of them, vinblastine and vincristine have been used as cancer chemotherapies. For the production of those chemical, a large amount of leaves are needed because the plant are only able to produce the chemical in small quantity. Therefore this research aims to investigate the effect of nutrient and irrigation level on the plant at different altitude, as an approach for a good agricultural practice in order to help the mass production of this plant. Early result found that the plant was grown larger at low altitude as the plant height, the number of leaves and flower was produced higher by plant in low altitude, but the leaf area was higher in plant at high altitude. The effects of water and nutrient treatments were varied in both sites. However, in general the high nutrient, but low ground water has a better effect on the plant growth. This early finding suggested that plant was growing better at lower altitude with high soil nutrients.

I. INTRODUCTION

Catharanthus roseus (L.) Don. or commonly also known as Madagascar periwinkle (Fig. 1) is a plant that has quite extensive been studied. The studies have emphasized on the chemical content as it’s used for natural medicines. This plant has been known because of commercially valued for sheltering more than 130 bioactive Terpenoid Indole Alkaloids (TIAs). Two of them that have been widely used for several anticancer chemotherapies are vinblastine and vincristine – the leaf-derived bisindol alkaloids (Chung et al. 2011; Man et al. 2012; Verma et al. 2012).

Chung et al. (2011) states that one well-studied plant metabolite is terpenoid indole alkaloids (TIAs) a class of alkaloids. A commercial source of TIAs such as vinblastin and vincristine were extracted from stems and leaves of Catharanthus roseus are used for anticancer drugs (Chung, et al., 2011). Other alkaloids such as ajmalicine and serpentine can be used to treat hypertension.

1

Figure 1. The Madagaskar periwinkle (Catharanthus rosues (Linn.) Don).

The information of the wealth of the C. roseus plants today have already available regarding their biosynthesis from plant organelles and tissues (Ferreres at al. 2011) or even up to the enzymes and corresponding genes levels (Facchini 2001, 2006; Facchini and De Luca 2008; Ziegler and Facchini 2008; Oudin et al. 2007). Recent studies on C. roseus plant have been conducted on the effect of bacteria for the generation of that plant (Wang et al. 2012) and the use of nanoparticle of C. roseus leaves as antiplasmodial activities (Ponarulselvam, 2012). Earlier studies found that the important C. roseus terpenoid for the pharmaceutical industry was due to the content of bisindole alkaloid vinblastine (Noble, 1990). It is currently used to treat a wide variety of neoplasms and is recommended for the treatment of Hodgkin’s disease, acute leukemia, and choriocarcinoma which is resistant to other therapies (Miura et al. 1987). However, the productivity of vinblastine is very low in plants (0.001– 0.0003%) (Uniyal et al. 2001) resulting in extraordinarily high price. Therefore, other studies in regard to improve the production of this valuable plant are necessary.

As the extensive studies that have been conducted on the chemical biosynthesis of the plants and recently the studies have been improve to the used of bacteria for the generation of C. roseus, to our knowledge it has been very rare studies conducted on the effect of environment, particularly the effect of altitude and nutrient and water supply for the plant growth and their production of plant chemical content where the plant grown in the field. However, the effect of slow increasing drought stress on plant has been found to increase of photosynthetic value in C. roseus (Kim and van Relsel, 2011). Few studies have also been

2 conducted on the effect of nitrogen supply on C. roseus, but the plants have been grown in a growth chamber (Ferreres at al. 2011; Kim and van Lersel 2011; Guo at al.2012). As for the application of GAP, the information of the environmental growth condition in the field for the C. roseus is very importance to evaluate due to its commercial and wealth chemical contents. Although the well known of its alkaloid as pharmaceutical values, the effect of soil where particularly the combination of water and nutrient levels to the plants growth as factors that may affect the production of plant chemical contents has not been well studied. Moreover, C. roseus can be found growing either in rural or urban area, the effect of the altitude to the growth and production of plant chemical contents also has not been assessed. This study aims to investigate the soil water and nutrient supply to the growth of C. roseus at different altitudes. The results will be useful for the farmers that would plant the C. roseus as for the application of GAP.

II. LITERATURE REVIEW

2.1. Catharanthus roseus

Catharanthus roseus (L.) Don., is a well-known medicinal tropical perennial sub- shrub belonging to the family Apocynaceae. This plant was indigenous to Madagascar and also known as the Madagaskar Periwingkle, now widely distributed, and is cultivated in China, India, Indonesia, Australia, North and South America (Yang et al. 2011; Verma et al. 2012). This is perennial shrubby plants, quite commonly found as a front garden plantation. It is originally from Central America and nowadays can be found in many tropical areas. In Indonesia it has many local names such as kembang tapak dara (Java/Indonesia), sindapor (Sulawesi), kembang tembaga (Sunda). In Malaysia it was known as kemunting cina, kembang sari cina, while in Philippine known as tsitsirika, in English known as periwingkle, in China known as chang chun hua, and in Netherland known a soldaten bloem (Tjitrosoepomo 1985; 1989).

They grow well from low to high altitude (up to 800 m above sea level) and prefer an open place, but it can also grow under canopy cover. The plant grow aside with many branches, 0.2 – 1.0 m tall. The leaves, petioles and twigs produce a milky exudate. The leaf blade abovate, green, and cross placed between the newer and the older growing leaves. The length of the leaves is between 20 – 60 mm, and 10 – 30 mm width, with short petioles. The branches and leaves covered by white latex. Flowers grow from axial with linier calyx lobes, the corolla lobes surrounded by white hairs with white, blue, pink or purple color depends on its cultivar. The length of the lobes is about 22 mm and the tube about 27 mm, with white

3 pollen. Carpels united by the style at the apex. The fruit/pod are cylindrical with pointy tips, with hair, 15 – 25 mm long, contain lots of seeds. Fruits may fall from the plant when they are still green, funicle attached in the middle of a saucer-shaped depression in the side of the seed. Testa rugose, embryo is green, with cotyledons slightly wider than the radicle (Tjitrosoepomo 1985; 1989).

2.2. Catharanthus roseus alkaloid content and function

Many studies have highlighted that this plant contain of about 130 Bioactive Terpenoid Indole Alkaloids (TIAs). Some of them have been intensively studied due to its ability to cure diseases such as vinblastine and vincristine (Chung at al. 2011; Verma et al. 2012; Man et al. 2012; Rakotoniriana et al. 2012; Wang et al. 2012). Other biosynthesis chemical content has also been studies was Vindoline, Catharanthine, Serpentine, Ajamlicine vinorelbine, vinflunine (Yang et al. 2011; Wang et al. 2012), and Catharoseumine (Wang et al. 2012).

The important of C. rosues plant as a source of medicinal plant has been widely known. Two Terpenoid Indole Alkaloids (TIAs), vinblastine and vincristine, are known as anticancer chemotherapies (Chung at al. 2011; Verma et al. 2012; Man et al. 2012; Wang et al. 2012). Catharanthus rosues plant has also been uncovered to produce a substance which acts as antifungal or antibiotic. The substance has been identified as a fungichromin, a known methylpentaene macrolide antibiotic, was the main antifungal component of TS3RO strain (Rakotoniriana et al. 2012). Ponarulselvam et al. (2012) found that the alkaloids produce by this plant are able to suppress the development of Plasmodium falciparum, a malaria parasite. The finding was also been proven that Plasmodium falciparum falcipain-2 was inhibited by one of the terpenoid indole alkaloids called Catharoseumine produced by C. rosues (Wang et al. 2012).

The most abundant substances found from the extract of this plant were Monomeric indole alkaloids, vindoline and catharanthine (3.8 and 7.7%, w/w, respectively) (Lopez et al. 2011). These chemical substances are used for the semisynthesis of 2 anticancer drugs, vinorelbine and vinflunine (Kruczynski and Hill, 2001; Montalar et al. 2011). Patient suffered from advanced human non-small-cell lung cancer and breast cancer is recommended to be treated with vinorelbine (Kanard et al. 2004). Vinflunine derived from a fluorinated analogue of vinorelbine, is being trialled for phase III bladder cancer treatment (Krzakowski et al. 2007).

4 2.3. Good Agricultural Practice

According to FAO the concept of Good Agricultural Practice (GAP) should meet the available knowledge to utilise the natural resources base in a sustainable way for the production of safe, healthy food and non-food agricultural products, to fulfil economic viability and social stability. In order to achieve these goals, comprehensive management strategy is required. The success will be depends on the developing skill and knowledge bases and the use of expert advised as needed. Furthermore, it explains that a good agricultural frame works may help to draw concern, discipline and practice therefore the individual production systems can be prepared within specific agro-ecosystems.

Agro-ecosystems aspects that play important roles are the physical and chemical structure and the biological activities of soil which can help for sustaining agricultural productivity, thus the soil fertility. Good soil management will help to maintain and improve soil fertility by minimizing losses of soil, nutrients, and agrochemicals through erosion, runoff and leaching. Good agricultural practice should manage farms in accordance with the properties, distribution, and potential uses of the soils, maintaining and improving soil organic matter by minimize erosion. Application of agrochemicals and organic/inorganic fertilizers in appropriate amounts and timing are important to agronomic and environmental requirements. Other aspect of GAP is water management in agriculture which takes a big responsible for water resources either in quantitative and qualitative terms. Cautious management of water resources and the efficiency of water use for crop production and irrigation should be included on the criteria. Other criteria include maximizing the infiltration of rain water on agricultural land and maintaining soil cover to avoid surface run-off and minimize leaching to water tables. The maintenance of adequate soil structure, including continuous macropores and soil organic matter, are important factors to achieve GAP. Efficient irrigation methods and technologies will minimize losses during the supply and distribution of irrigation water by adapting the quantity and timing to agronomic requirements to avoid excessive leaching and salinization. Therefore, the water tables should be managed to prevent excessive rise or fall. Good agricultural practice will avoid water contamination by maximising water infiltration and minimising unproductive efflux of surface waters from watersheds, manage ground and soil water. This can be done by building up soil structure and soil organic matter, apply production inputs, including waste or recycled products of organic, inorganic and synthetic nature. Adopting techniques to monitor crop and soil water status, accurately

5 schedule irrigation, and prevent soil salinization are important by adopting water-saving and re-cycling. Increasing the function of the water cycle by establishing permanent cover, or maintaining and restoring water bodies are also important. Type of crops, their cultivars and varieties chosen to meet local consumer and market need should be met their suitability to the site. Perennial crops are used to provide long-term production options and opportunities for intercropping. Annual crops may grow in sequences, including those with pasture, to maximize the biological benefits of interactions between species and to maintain productivity. Harvesting of all crop and animal products removes their nutrient content from the site and must ultimately be replaced to maintain long-term productivity. Maintaining crop health is essential for successful farming for both yield and quality. This requires long-term strategies to manage risks by the use of disease- and pest-resistant crops, crop and pasture rotations, disease breaks for susceptible crops, and the minimal use of agrochemicals to control weeds, pests, and diseases following the principles of Integrated Pest Management. The quality of the product depends upon the protocols for harvesting, storage, and processing of farm products. Harvesting must conform to regulations relating to pre-harvest intervals for agrochemicals. The products should be stored under appropriate conditions of temperature and humidity in space designed and reserved.

III. MATERIAL AND METHOD

3.1. Location and Time of study

The research has been conducted in two locations with two different altitudes, Renon, Denpasar (Fig. 2a) with the altitude of <10m above sea levels, and Pancasari, Tabanan with altitude of 800m above sea levels (Fig. 2b). Seedling preparation had been started from June 2012, and planted from July 2012 for 3 moths before the seedlings were transferred to the field. Field works were started from September to Desember 2012.

Material used for this research are:

1. Seedling preparation:

- Seed of C. roseus

- Cups and small poly-bag to plant the seed

- Soil and caw manure

6 - Hose

2. For the plant treatments and harvesting

- seedlings - ruler

- Fertilizers (ZA) - Bamboo steaks

- hose - Scissor

- buckets - Plastic bags

- shovel - Paper marks

- tape measurement - Trays

a b

Figure 2. The Madagaskar periwinkle grow at Renon, Denpasar (a) and Pancasari, Tabanan (b).

3.2. Seedling preparation

Seed of C. roseus were collected from Denpasar then grown in Sanur (about 3km from Renon). Good seeds were selected by adding the seeds with water, and then the drowning seed were selected to be grown. Few seeds were placed in a small plastic cups. Seedling started to grow after 1.5moth, then after the seedling were 3 months old, they transferred to the field sites (Fig. 3).

7

Figure 3. Three months old seedlings of Madagaskar periwinkle (C. rosues).

3.3. Field Works: Planting and Caring

Before growing the plant in the field, soil was collected from the two sites (Renon, and Pancasari) to be nutritionally analysed. Samples were sent to Soil Laboratory, Faculty of Agriculture University of Udayana. The soil minerals analysed were pH, Carbon, Phosphor, Kalium, soil water content, soil electric conductivity, and soil texture.

Three moths old seedling were transported to the field sites to be planted. Land in each field location was selected based on the availability. A 150x100m piece of land was used for each site. Land was prepared by shovelling and mixing the soil. The land was divided into 9 parts that was split by a small ditch. Each part of land was treated with different water and nutrient treatments or its combinations. For each treatment 12 individual seedlings were planted.

Figure 4. Planting and caring the Madagascar periwinkle (C. roseus) before the water and nutrient treatments.

8 All planted seedling was watered daily until the plant grow well before different the water and nutrient treatments were carried out. The water treatment was performed after 2 weeks, and the nutrient treatment was performed after 1 month of planted seedlings.

Figure 5. The Madagaskar periwinkle before nutrient and water treatment was performed at Renon, Denpasar.

Figure 6. The Madagaskar periwinkle before nutrient and water treatment was performed at Pancasari, Tabanan.

3.4. Plant water and nutrient treatments

Each portion of land that has been planted with C. roseus plant treated with different water and nutrient treatments. The water treatments were divided into 3 categories: control (plant was watered if necessarily), low (plant was watered with 100ml daily), and high water treatments (plant was watered with 200ml twice a day). The nutrient treatment was only

9 applied once after the plant was 1 month old, which divided into control (no nutrient addition), low nutrient (each plant treatment was added with 3g of ZA fertilizer) and high nutrient treatment (each plant treatment was added with 6g of ZA fertilizer). The combination of the two treatments (water and nutrient) was also conducted. The plant growth i.e the plant height (the tallest branch), leaf area of 10 leaves measured from the third grown leaves, number of branches and the number of open flowers, was measured weekly.

Figure 7. The Madagascar periwinkle after water and nutrient treatment was performed for 4 weeks at Renon, Denpasar.

Figure 8. The Madagascar periwinkle after water and nutrient treatment was performed for 4 weeks at Pancasari, Tabanan.

10 3.4. Plant Harvest and Lab Works

Plants were harvested by pulling the whole individual plants after 6 weeks of the nutrient treatments. Plants were sorted according to the treatments. After harvesting the plants were transported to and dismantle. Each of plant part was separated and counted i.e. the number of branches, number of flower, number of leaves and fruits per branch, leaf area, leaf mass, and whole plant above and below ground biomass (Fig. 9a). Leaves were air-dried for the purpose of plant chemical analyses (Fig. 9b).

a b

Figure 9. Harvesting C. roseus plants (a) and air-dried leaves (b).

The assessment of plant biomass was conducted by measuring the wet weight of the below ground (roots) and above ground (braches and leaves) plant material. Those plant materials then were oven dried at 70°C until constant weight at ‘Ecology Laboratory and Plant and Physiology Laboratory, Department of Biology, Faculty of Mathematic and Science, University of Udayana. The plant chemical contents such as Nitrogen and its metabolites were assessed from the air-dried leaves. The air-dried leaves then finely ground using a blender. The grounded leaf materials then send to the ‘Laboratorium Nutrisi, Faculty of Agricultural Technology’ and in ‘Laboratorium Analitik, University of Udayana for the analyses. Up to date, the plant chemical contents are still being analysed.

IV. RESULT AND DISCUSSION

4.1. Soil Type and Mineral Content

Type of soil from the two sites (Renon and Pancasari) was quite different. The soil water content was twice higher at Pancasari (7.18%) than at Renon (3.85 %). The type of soil at Renon was sandy soil but clay at Pancasari. Similarly the soil pH was not similar, which

11 was slightly acid (pH = 5.9) at Renon and neutral (pH = 6.9) at Pancasari. The soil nitrogen content was low at both sites (0.15 ppm at Renon and 0.19 ppm at Pancasari), but the soil phosphate concentration was very high at Renon (363.68 ppm) and high at Pancasari (15.39ppm), while the soil potassium concentration was good at Renon (151.51 ppm) but low (116.94 ppm) at Pancasari. The soil electrical conductivity was low at Renon (1.01 mmhos/cm), but high in Pancasari (3.14 mmhos/cm). It has morphologically shown as in Fig. 10 that C. roseus were able to grow quite well in both sites, suggesting that those plant was adapted to both site.

a b

Figure 10. The growing Madagascar periwinkle (C. roseus) in low altitude at Renon (a) and high altitude at Pancasari (b) before treated with different water and nutrient treatments.

4.2. The growth preference of C. roseus plant at different altitude

The results of this research show that the plants can be grown well in both low to high altitude. However, plants growing in lower altitude grow better than plant in high altitude. This has visually recognised from the size of the plants (Fig. 11). The plants grow in lower altitude has larger size than plants grow in high altitude. They are also taller in low altitude than in high altitude.

12 a b

Figure 11. The C. roseus grows at Renon (a) where the individual plant was larger and taller than that at Pancasari (b).

The plant growth was different between plants grow in low and high altitude (pooled data, regardless of treatments). The plants in lower altitude grow higher than those in high altitude. This can be seen that the plant height in lower altitude (Renon) was twice higher than those in high altitude (Pancasari) (Figg. 12). The finding was corresponded with the study by Kofidid et al. (2003) that the increase in elevation result in a progressive decrease in plant height.

Renon 400 Pancasari 350

300

250

200

150

Average per plant Average 100

50

0 Height (cm) No. leaves No. Flower Leaf Area (mm)

Figure 12. Average plant height, number of leaves, and number of flower of C. roseus grow in low (Renon) and high (Pancasari) altitudes.

The number of leaves per plant was also higher on plants grow in lower altitude than plants grow in higher altitude, but the number of flower was similar. Although plants grow at lower altitude (Renon) produced higher number of leaves, they produced smaller size of leaves. A further analyse for the leaf mass should be conducted to predict the plant production.

13 4.3. The growth rate of C. roseus plant at different water and nutrient treatments

Plant growth rates of each water and nutrient treatments was varied between and within treatments. Plants grow at lower altitude (Renon) showed that the plant height was similar among all nutrient and water treatments (Fig. 13). It seem that fertilizer have a slight effect to the number of leaves and number of flowers per plant. Plants treated with high fertilizer grow more leaves and flowers than plant with low or no fertilizer (control). The results shows that nutrient addition to the soil has a positive effect to the plant grown since the area used for planting contain low nitrogen. It has been widely recognised that nitrogen is important for the plant growth (Uchida 2000; de Graff et al. 2006).

The water treatment was not seems to have an effect the plant growth. However, plant treated with higher water seems to grow slower by having less number of leaves than control and plant treated with less water (Fig. 13). This result may indicate that this plant is better to grow in drier habitat than in wet habitat, as the C. roseus plant is a perennial shrubby plant that mainly found in tropical area.

500 Height (cm) # leaves 400 # Flowers

300

200

Average per Plant Average 100

0 C HF LF HW LW Treatments

Figure 13. Average plant height, number of leaves and number of flower of C. rosues grow in low altitude (Renon) from different water and nutrient treatments. Note: C = Control; HW = High Water; LW = Low Water; HF = High Fertilizer; LF = Low Fertilizer.

Plant treated with different water and nutrient treatments grown in higher altitude (Pancasari) showed that there was no variation in plant height, but as plant grown at lower altitude, the number of leaves was slightly higher on plant treated with more fertilizer. The number of flowers, however, was double produced by the plant in high fertilization (Fig. 14).

14 Height (cm) 180 # leaves 150 # flowers

120

90

60 Average per Plant Average 30

0 C HF LF HW LW Treatments

Figure 14. Average plant height, number of leaves and number of flower of C. rosues grow in high altitude (Pancasari) from different water and nutrient treatments. Note: C = Control; HW = High Water; LW = Low Water; HF = High Fertilizer; LF = Low Fertilizer.

The leaf area of C. roseus plant at lower altitude was not affected by nutrient and water treatments. On the other hand, the leaf area of plant grow in higher altitude (Pancasari) was varied among treatments (Fig. 15). Interestingly, the leaf area was higher in control treatment than those with low or high nutrient and water treatments. This results did not sustain the finding by Kim and van Lersel (2011) that the dry condition affect the physiological responds of this plant by producing higher total leaf are per plant. However, further analyses for the other parameter such as the leaf mass should be taken to prove the findings.

Pancasari 40 Renon

30

20

10 Av. Leaf Area (mm) Area Av. Leaf

0 C LW HW LF HF Treatment

Figure 15. Average leaf area of C. rosues grow in low altitude (Renon) from different water and nutrient treatments. Note: C = Control; HW = High Water; LW = Low Water; HF = High Fertilizer; LF = Low Fertilizer.

15 4.4. Good Agricultural Practice of Catharanthus roseus

The early results of this project for the Good Agricultural Practice of the Madagascar periwinkle (C. rosues) suggest that this plant in general was better growing in lower altitude than in higher altitude. As can be seen from the results that C. roseus grow in low altitude (<10 m above the sea level) at Renon was better and faster than those at Pancasari with the altitude of 800 above sea levels. It has also been studied that the main part of plants that are useful for the production of plant metabolites are the leaves. This mean that the plant may be better to grow in lower altitude, as they produce more leaves that those at higher altitude. However, it should be noted that the number of leaves produced by plant was not the only character that affect the production of plant metabolites, but the leaf area or leaf mass has been known to affect the production of plant metabolites (Sun 2006; Watiniasih 2009).

It has been mention before that up to date the recommendation has just based on a small number of data. The results that have been collected were merely from the plant growth rate of each treatment and the preceding soil type and chemical contents. Further analyses of, for example, the leaf chemical contents and plant biomass will be very important. Base on the current results for the Good Agricultural Practice and for the mass production of the Catharanthus roseus plant is recommended that the plant is better to cultivate in lower altitude, such as at Renon.

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Acknowledgement

We thank the PIC-TPC Unud Project for the financial support. We also grateful to some Biology students: Saka, Dyah, Melandani, Royana, Timothy, Wahyu, and Buya for their help on harvesting the plants. We are indebted to the family of Bapak …. and Bapak Made Wirata for providing the land and help.

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