VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE FACULTY OF BIOTECHNOLOGY ------------------------------ GRADUATE THESIS TESTING THE EFFECT OF KAEMPFERIA GALANGA L. ON THE LARVAE OF PIERIS RAPAE L. HANOI-2021 VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE FACULTY OF BIOTECHNOLOGY ------------------------------ GRADUATE THESIS TESTING THE EFFECT OF KAEMPFERIA GALANGA L. ON THE LARVAE OF PIERIS RAPAE L.
Student : Trinh Thi Huong Tra ID : 610681 Class : K61CNSHE Supervisor : Dr. Dang Thi Thanh Tam HA NOI-2021 COMMITMENT I hereby declare that all information in this document has been obtained and presented in accordance with academic rules and ethical conduct. I also declare that all the help to complete this thesis has been thanks. Trinh Thi Huong Tra i ACKNOWLEDGEMENT This study would not have been possible without the support of many people.
I would like to express my gratitude to my professors of Department of Plant Biotechnology at Faculty of Biotechnology in Vietnam National University of Agriculture for their support and conducting the research. Particular thanks to Dr. Dang Thi Thanh Tam for her valuable comments and helpful advice, scientific , and valuable feedback throughout the time studying at the laboratory of the Department of Plant technology. I would also like to extend my partner Nguyen Dinh Thinh for helping me to perform some experiments.
Finally, I would like to express my special thanks to my family for great encouragement as well as my friends for their supports. Trinh Thi Huong Tra ii CONTENTS COMMITMENT. iii LIST OF TABLES. v LIST OF FIGURES.
Purpose and requirement. Some researchs on Pieris rapae. MATERIALS AND METHODS. Instruments and chemicals.
Pieris rapae culture. Preparation the plant extracts. 19 iii PART IV. RESULTS AND DISCUSSIONS.
CONCLUSIONS AND SUGGESTIONS. 28 iv LIST OF TABLES Table 4. Mortality rate of larvae during Kaempferia galanga L.2 Dose–response parameters of Pieris rapae after 24 hours. Deterrence percentage acreage leaves of Kaempferia galanga extract on Pieris rapae larvae.
24 v LIST OF FIGURES Figure 2.1 Butterfly and larvae of Pieris rapae .2 Eggs of Pieris rapae .3 Five stages of Pieris rapae larvae .4 Pupae of Pieris rapae .5 Adult of Pieris rapae .6 Kaempferia galanga plants .2 The process of growing cabbage .3 Pieris rapae culture .4 Preperation steps of the Kaempferia galanga L extract .5 Steps of toxicity tests .6 Steps of antifeeding test. Mortality rate of larvae after 24 hours of treatment .2 Testing toxicity of (4%) Kaempferia galanga extract on Pieris rapae larvae.3 The leaf consumption of larvae in the treatment with 0.5% Kaempferia galanga extract .4 The leaf consumption of larvae in the treatment with 1% Kaempferia galanga extract .5 The effect of Kaempferia galanga extract on the antifeeding of Pieris rapae larvae. Introduction In Vietnam, cruciferous vegetables (broccoli, cabbage, cauliflower, kale, collards .) are preferred by many people because of their rich nutrients. They have high economic value and are widely grown.
Cruciferous vegetables are the main crops of many regions such as Sapa, Son La, Đa Lat,.However, cruciferous vegetables have a short growth time, so they are planted continuously, harvested in scattered batches. Besides, they have the features of stems, soft and weak leaves, containing many nutrients. It is suitable with the warm and humid climate of our country, so it is damaged by many kinds of worms such as silkworm, Pieris rapae, click beetles ., seriously affecting the productivity and quality of vegetables (Trinh, 1999). In the cruciferous vegetable pest group, Pieris rapae is the most dangerous pest for vegetables.
Every year they appear from 14-15 times, with high density (Lam, 2008). Pieris rapae is a small to medium-sized butterfly species of the family Pieridae. It is believed to have originated in the Eastern Mediterranean and widespread in Europe and Asia. The butterfly is recognizable by its white color with small black dots on its wings.
Young caterpillars gnaw the green matter of vegetable leaves. From stage two or more, they gnaw leaf blades and remain only the veins. Their waste can spoil the leaves. So if the high density is left, the field of vegetables will be bare and frayed.
To control Pieris rapae, the farmers mainly use chemical methods. In vegetable-growing areas, pesticides are used extensively and continuously. Chemical control is highly effective but it contains negative risks to organisms, the ecosystem and human health. These drawbacks associated have spurred the search for biological pesticides.
Because biological control is an environmentally friendly method for the environment. In this field, plant 1 extract is a solution commonly. However, this method still exist some limitations. It is more susceptible to environmental conditions, slow action and sometimes is unpredictable.
Because of the reasons above, the study was performed to find the plant extracts that have highly effective insecticide on Pieris rapae. Purpose and requirement 1. Purpose Testing the effect of Kaempferia galanga extract to control Pieris rapae. Requirement Testing the toxicity of Kaempferia galanga extracts on young caterpillars.
Testing the antifeeding effect of Kaempferia galanga extracts on young caterpillars. Research location The study was performed at Department of Plant Biotechnology ,Faculty of Biotechnology and Institute of Agro- Biology in Vietnam Nation University of Agriculture. Pieris rapae Classification Order Lepidoptera Family Pieridae Genus Pieris Species P. Rapae Binomial name Pieris rapae Figure 2.1 Butterfly and larvae of Pieris rapae Pieris rapae is a small to medium sized butterfly species of family Pieridae.
The butterfly is recognizable by its white color with small black dots on its wings. Their hot plants are crucifer crops such as cabbage, kale, bok choy and broccoli…. Pieris rapae is found principally in temperate regions of Europe, Africa, Asia, and North America, but now it appear in Australia and New Zealand, southerm Mexico, and Hawaii… Imported Pieris rapae was first observed in North America in 1860 (Beirne, 1971). The life cycle of Pieris rapae has four stage: egg, larvae, pupae and butterfly.
It requires three to six weeks and depending on weather and temperature. Imported cabbageworm can be found throughout the year in Vietnam and the most quantity at time from autum to winter when the source food is plentiful.2 Eggs of Pieris rapae The egg is yellow and has surface ridges running lengthwise and crosswise. The egg is laid on end, usually on the under surface of the leaves. They occur singly, never in clusters.
Eggs hatch in 3 to 7 days (Ronald, 2007). Larvae : The green caterpillar has a slender yellow stripe along the middle of the back, and another intermittent yellow stripe along each side of the body.The instar have five stages. The body length are about 0. Body lengths at maturity of each instar averages from 2 to 3 cm respectively.
The larvae requires about 15 days (range 11 to 33 days) to complete its development. The best condition to develop at 20-24°C temperature and 75% - 85% humidity (Muggeridge,1942).3 Five stages of Pieris rapae larvae The 1st instar of Pieris rapae after hatching, the body of the larvae is ivory-white or milky-white, sometimes pale yellow. Average length of 1st instar lavae body is 2.043mm, average body width is 0. At 2nd instar, the lavae has a green color similar to the color of the leaf, the body has more feathers.
The average length is 6. Body width ranges from 0. The body of the 3rd instar is green, the head is light brown. Average body length is 13.
Average body width is 0. The 4th instar of Pieris rapae has the maximum number of fluff. Average body length is 19. Average body width is 0.
Larvae at stage 5 is very voracious, maximum body size, the body still is green and covered with fluff. Body length ranges from 25. Body width ranges from 1. 5 Pupae : Pupae is usually green, but gray or tan colored pupae occur often.
It has sharp angular projections over the back and front. Pupae are attached to the under surface of leaves by silk at the tail and a silk rope near the midsection. Pupae are about 3/4 inch long and are found on the plant or in a sheltered area up to 15 yards away from the host plant. Pupal development is completed in 1 to 2 weeks (Ronald, 2007).4 Pupae of Pieris rapae Adult The butterfly has a wing span of about 1-1/2 inches.
The forewings are black-tipped. A female has two black spots on top of each of her forewings; a male has only one spot (Ronald, 2007). When viewed from below, the wings generally are yellowish, and the black spots usually show faintly through the wings. The body of the butterfly is covered with dense hair, which is colored white in females, but darker in males.
The adult typically lives about three weeks. The female produces 300 to 400 eggs. The adult is very active during the daylight hours, often moving from the crop to flowering weeds to feed (Harcourt,1962).5 Adult of Pieris rapae The Pieris rapae larvae is voracious. Once it hatches from the egg, it eats its own eggshell and then moves to eat the leaves of the host plant.
It bores into the interior of the cabbage, feeding on the new sprouts. The mustard oil from the host plant makes the larvae distasteful to birds. The larvae adjust their feeding rate to maintain a constant rate of nitrogen uptake. They will feed faster in low nitrogen environment and utilize the nitrogen more efficiently (at the cost of efficiency in other nutrients) than larvae hatched on high nitrogen host plant.
However, there is no significant difference in growth rate was observed between larvae in the two environments ( Scott, 1986). Considered a serious pest, the caterpillar is known to be responsible for annual damage worth hundreds of thousands of dollars (Holland, 1931). The larvae are shown to disperse their damage on the plants. Larvae are shown to feed mostly during the day.
They move around the plant mostly spending their time feeding. A feeding bout is immediately followed by a change in position, either to a new leaf or to another part of the same leaf (Maurico, 1990). This dispersal of damage is seen as an adaptive behavior to hide the visual cues from predators that rely on vision. Even though Pieris rapae larvae are cryptic, they remain in the sun for the majority of the day, rather than hiding on the underside of the leaf.
The condition of the host plant influences the larvae growth significantly. From the stage two or more, they 7 gnaw leaf blade and remain only the veins. Their waste can spoil the leaves. So if the high density is left, the field of vegetables will be bare and frayed.
Some researchs on Pieris rapae For the past 30 years, scientists have been studying about plant extracts and phytochemicals to develop the insecticides that reduce the human health and environment. The damage due to insects may be up to 10–20% every year (Ferry et al. To reduce the damage caused by Pieris rapae on cruciferous vegetables, many authors from over the world have been had many studies on different kinds of plant extracts and had obtained positive results. Seyedeh et al ( 2011) performed a study on Pieris rapae with two medicinal plants ( Artemisia annua L and Achillea millefolium L.
The Artemisia annua and Achillea millefolium extract were extracted on methanol slovent by procedure of Warthen et al (1984). Toxicity tests were performed at 0.5%, 5%, and 10% concentrations of the extracts on larvae at stage 3 of Pieris rapae. It was followed after 24h and 48h and determined LC5, LC25. The result showed that after 48h LC50 and LC25 values of Artemisia annua L were 9.19%, LC50 and LC25 values of Achillea millefolium L were 3.
Achillea millefolium L extract was more toxic. Deterrency tests were the tests to determine the leaf consumption of young caterpillars. The treatment leaves were treated at 0.