Infection of Phytophthora Palmivora From Soil in Cocoa Plantation

Phytophthora palmivora causes serious losses on cocoa in Indonesia and world-wide. The research aimed to assess the potential of soil as source of inocula for Phytophthora diseases in cocoa. Soil samples were baited using a healthy cocoa pod tissue, and the pathogen was isolated for morphological and molecular identification. Baiting technique was successfully used to detect the presence of P. palmivora in soil samples, and this was confirmed by morphological and molecular identification. P. palmivora can be detected in soil in all year around in wet areas indicating that soil is a massive and consistent source of inocula. Surveys conducted on the soil of Amazonian, Amelonado and Trinitario blocks of various ages showed that P. palmivora can be found in old and young cocoa blocks, even as young as 3 or 4 years. P. palmivora infection from soil to the pods appears to be mainly through contact or rain splash. Baiting with whole healthy pods exposed at different heights above undisturbed litter and above bare soil showed that the infection still occurred at 100 cm above the soil, even though it decreased gradually with the height. Infection from litter was not different to that from bare soil, indicating that the litter layer is not acting physically as a shield preventing rain from splashing the inocula up from wet soil to the pods. However, in tests for the possibility of P. palmivora carried through air convection, no pod was found to be infected, suggesting that the pathogen was not carried through convective accend of aerosol droplets from soil surface up to pods in the canopy

PERUBAHAN BIOLOGIS DAN FISIOLOGIS SEBAGAI INDIKATOR MASAK BENIH KAKAO HIBRIDA

ABSTRAKProgram pengembangan dan rehabilitasi tanaman kakao membutuh-kan benih bermutu. Mutu benih antara lain ditentukan oleh saat panenyang tepat, terutama berhubungan dengan masak fisiologis. Beberapaindikator penting yang berkaitan dengan masak fisiologis benih adalahkarakteristik biologis dan fisiologis. Penelitian telah dilaksanakan diKebun Induk Benih Pusat Penelitian Kopi dan Kakao (Puslitkoka) Jember,Laboratorium Fisika dan Laboratorium Ilmu dan Teknologi Benih IPB,serta Rumah Kaca Balai Penelitan Bioteknologi Perkebunan Indonesia,Bogor pada bulan Februari-September 2008. Penelitian ini bertujuan untuk(1) mempelajari perubahan biologis dan fisiologis selama perkembanganbenih kakao hibrida, (2) mengetahui hubungan antar berbagai karakterbiologis dan fisiologis benih yang mencerminkan mutu benih, dan (3)menentukan saat panen yang tepat benih kakao hibrida TSH 858 xSca 6 dan ICS 60 x Sca 6. Benih yang digunakan berasal dari hasilpersilangan buatan antara kakao TSH 858 x Sca 6 dan ICS 60 x Sca 6.Umur panen benih yang digunakan dalam penelitian adalah 120, 135, 150,165, dan 180 hari yang dihitung saat setelah antesis, dan setiappengamatan diulang 4 kali. Analisis data disajikan dalam bentuk grafikdengan data primer ditambah standar deviasi dalam program Excel danuntuk mengetahui hubungan dari masing-masing karakter mutu benihdilakukan ”analisis path” menggunakan SAS dari Windows v 9.1. Hasilpenelitian menunjukkan dua fase perkembangan benih. Fase perkem-bangan hingga masak fisiologis (fase 1) dan fase setelah masak fisiologis(fase 2) kakao hibrida TSH 858 x Sca 6 dan ICS 60 x Sca 6. Masakfisiologis benih kakao hibrida TSH 858 x Sca 6 tercapai pada saat 150HSA dan ICS 60 x Sca 6 pada 165 HSA. Daya kecambah, indeks vigor,K CT -R, T 50 , bobot basah dan bobot kering benih, karotenoid dan antosianinbenih dan buah, jumlah daun, dan tinggi bibit dari benih kakao hibridaTSH 858 x Sca 6 dan ICS 60 x Sca 6 mencapai maksimum pada saatmasak fisiologis dan menurun pada fase kedua. Selama periodeperkembangan benih terjadi penurunan total klorofil benih dan buah,sedangkan warna buah kuning mengalami peningkatan. Karakter yangberhubungan langsung dengan mutu benih pada saat masak fisiologisbenih kakao hibrida TSH 858 x Sca 6 dan ICS 60 x Sca 6 adalah warnabuah kuning, indeks vigor, total klorofil benih dan buah, karotenoid danantosianin benih, T 50 , tinggi bibit, K CT -R, dan bobot kering benih.Kata kunci: Theobroma cacao, biologi benih, fisiologi benih, karakteristikbenih, mutu benihABSTRACTBiological and Physiological Changes as Indicator ofMaturity of Hybrid Cacao SeedThe development and rehabilitation programs of cacao need highquality seeds. The high quality of cacao seeds is influenced by seedsphysiological maturity and harvesting time. Several important indicatorsrelated to the seed physiological maturity are biological and physiologicalcharacters. The research objectives were: (1) to study biological andphysiological changes during of seed development, (2) to study on thecorrelation of various characteristics related with seeds physiology andquality, (3) to determine the most appropriate harvesting time for hybridcacao seed of TSH 858 x Sca 6 and ICS 60 x Sca 6. The research wasconducted at Coffee and Cacao Research Institute of Indonesia(Puslitkoka) in Jember, IPB Biophysics and Seeds and TechnologyLaboratory and Biotechnology Research Institute for Estate CropsIndonesia glass house in Bogor from February to September 2008. Theseeds were originated from hand pollination of TSH 858 vs Sca 6 and ICS60 vs Sca 6 hybrids from Puslitkoka Jember. The seeds for this researchwere harvested on: 120, 135, 150, 165, and 180 days after anthesis (DAA);with four replications each. Data were analyzed and presented as graphs,standard deviation in excel; while the relationship of each character ofseeds quality was determined using path analysis by SAS for Windows v.9.1. The results showed that the seed physiological changed on two phasesduring its development. The first phase started from seeds development upto physiological maturity for TSH 858 x Sca 6 and as well ICS 60 x Sca 6hybrids, and second phases started after physiological maturity. Thephysiological maturity of each seeds is 150 DAA for TSH 858 x Sca 6 and165 DAA for ICS 60 x Sca 6 hybrids. Seed germination percentage, vigorindex, germination rate (K CT -R and T 50 ), wet and dry weight of seed, seedsand fruits carotenoid content, seed and fruit anthocyanin content, numberof leaves, and height of seedling reached maximum when seed achievedphysiological maturity and decreased afterward. During seed development,there was decreasing of seeds and fruits chlorophyll content and increasedfor the yellow color of fruit. The characters which showed directcorrelation with seeds quality during seed development of TSH 858 x Sca6 and ICS 60 x Sca 6 hybrids are: yellow color of fruit, vigor index,chlorophyll content for seeds and fruit, seed carotenoid and anthocyanincontent, germination rate (T 50, K CT -R), seedling height and seed dry weight.Key words: Theobroma cacao, seed biological, seed physiological, seedcharacteristic, seed qualit

Chitinase and Peroxidase Activities, Stomatal Density and Resistance of CocoaAgainst Black Pod Disease

Research based on morphology which is related to structural resistance such as stomata density and chemical resistance involving chitinase and peroxidase enzymes needs to be conducted. Hereafter, cacao resistant mechanism against Phytophthora palmivora can be realized. Research took place in June 2008 till February 2009 in Plant Disease Laboratory and Experiment Station of Kaliwining, Indonesian Coffee and Cacao Research Institute, Jember, East Java. Research on chitinase and perxidase analysis took place in Inter University Center Laboratory of Bogor Agriculture University (IPB). Based on stomata observation of 10 clones, the stomata density in pod and leaf did not give high correlation to the resistance. Resistant cacao clone did not always low stomata density compared to susceptible ones. Cacao clone which was susceptible did not always have high stomata density in pod and leaf. The number of stomata did not give significant difference between resistant clones and susceptible ones. The activities of chitinase and peroxidase enzymes upon tested clones indicated that there was chitinase role in the resistance of cacao against the infection by P. palmivora fungi. The increase of chitinase activity fungus in resistant clones generally intensified consistently, and similiar with peroxidase enzyme

Sonogram Pemeriksaan Kebuntingan Dini pada Kambing Kacang (Capra Hircus)

This study was conducted to determine the earliest day of pregnancy diagnosis in kacang goat using transrectal ultrasonography. The goat were synchronized by using prostaglandin in the luteal phase. Pregnancy was determined by isoechogenic visualization surrounded by hypoechogenic. Early pregnancy was detected on days 20 of embryonic vesicle diameter 1.2±0.1 cm. Fetus was detected on days 22 with a long gestation fetus 0.4±0.1 cm. Average increase until days 30 pregnancy was 0.19 ± 0.1 cm per day. Development of fetus was followed by an increasing the diameter and thickness of uterus. The diameter of uterus increased from days 14 (0.8 ± 0.3 cm) until days 30 (3.6 ± 0.2 cm), and thickness of uterus increased from days 14 (0.4 ± 0, 2 cm) until days 30 (1.8 ± 0.2 cm). It could be concluded that the earliest pregnancy diagnosis showed positive sign on days 20 and fetus was earliest observed on days 22

Rapid Detection of the Africanized Honey Bee: a Tool for Indonesian Animal Quarantine

Molecular detection methods were used to determine if Africanized Honey Bees (AHBs) are present in populations of imported Apis mellifera in Indonesia. The cytochrome b (cyt b gene) was amplified from mitochondrial DNA and digested with the BglII restriction enzyme (cytb/BglII). Two types of animal DNA extraction kits were used and found suitable for rapid preparation of DNA from A. mellifera by the Animal Quarantine facility. Results showed that all 94 colony samples from beekeepers in Java produced a 485 bp PCR product from the amplification of this gene. Two DNA fragments of 194 and 291 bp from all samples were produced after digestion with BglII. This cytb/BglII result together with the DNA sequence of cyt b showed that all collected samples of A. mellifera were the non-AHB type. Hence, this study did not detect AHB in Indonesia

PATOGENISITAS BEBERAPA ISOLAT CENDAWAN TERBAWA BENIH KAKAO HIBRIDA

ABSTRAKBenih kakao hibrida diketahui dapat membawa beberapa mikrobayang bersifat patogenik dan menurunkan mutu benih. Penelitian bertujuanuntuk mengetahui pengaruh beberapa isolat cendawan terbawa benihterhadap penurunan viabilitas benih dan vigor bibit kakao hibrida.Penelitian dilakukan di Kebun Benih Pusat Penelitian Kopi dan KakaoIndonesia, Jember, Laboratorium Mikrobiologi dan rumah kaca BalaiPenelitian Bioteknologi Perkebunan Indonesia, Bogor, pada bulan Julisampai November 2008. Penelitian menggunakan 13 cendawan terbawabenih kakao hibrida. Benih diperoleh dari persilangan buatan antara kakaoTSH 858 dengan Sca 6. Penelitian menggunakan model Rancangan AcakLengkap dengan 4 ulangan. Inokulasi patogen pada benih kakao dilakukandengan cara merendam benih di dalam suspensi patogen dengan kerapatan10 6 spora/ml selama 30 menit. Selanjutnya benih ditanam pada media pasirsteril dalam boks plastik ukuran 30 x 30 cm, menurut rancangannya.Setiap perlakuan diulang 4 kali. Parameter yang diamati adalah dayaberkecambah, indeks vigor, kecepatan tumbuh relatif, kecepatanberkecambah T 50 , laju pertumbuhan kecambah, jumlah daun, tinggi bibit,panjang akar, jumlah akar dan kematian benih. Data dianalisis denganANOVA dan dilanjutkan dengan uji Jarak Berganda Duncan. Hasilpenelitian menunjukkan bahwa ke-13 spesies cendawan bersifat patogenikpada benih kakao hibrida. Cendawan patogen terbawa benih yang bersifatpatogenik adalah Aspergillus flavus, A. ochraceus, Cladosporiumherbanum,  Curvularia  geniculata,  Fusarium  oxysporum,  Phomaglomerata dan Macrophoma sp. Cendawan patogen tersebut dapatmenurunkan daya berkecambah 20-40%, indeks vigor 30-47%, kecepatantumbuh relatif 13-45%, dan meningkatkan kecepatan perkecambahan(T 50 menurun) dari 0,62-7,36 hari. Ke-13 isolat patogen dapatmenyebabkan kematian benih 29-52% dibanding kontrol. Ke-13 isolatpatogen juga menginfeksi bagian tanaman seperti kotiledon, daun, batangdan akar bibit kakao, namun hanya Phoma glomerata dan Macrophomasp. yang menurunkan tinggi bibit, jumlah daun, jumlah dan panjang akarsecara nyata. Tujuh dari 13 isolat cendawan patogen terbawa benih tidakhanya menurunkan viabilitas dan vigor benih kakao hibrida tetapi jugadapat berkembang pada bibit sehingga perlu penanganan benih secara dini.Kata kunci: benih hibrida, patogen terbawa benih, viabilitas, vigor benih,Theobroma cacaoABSTRACTIn 2009 revitalization of cacao plantations in Indonesia required 168million seeds. Distribution of low quality and infected seeds leads to hugelosses and in a long term will destruct cultivation of cacao. Seed-bornepathogens of infected cacao hybrid seeds are dangerous because they mayreduce physiological qualities of the seeds. The study aimed atdetermining the effect of several isolates of seed borne fungi on theviability and vigor of hybrid cacao seeds as well as growth of theseedlings. The study was conducted at the Seed Garden Indonesian Coffeeand Cacao Research Center in Jember, Microbiology Laboratory and glasshouse of Biotechnology Research Institute for Estate Crops of Indonesia,Bogor, from July to November 2008. The study used 13 seed-borne fungiin hybrid cacao. The cacao seeds were obtained from hand pollinatedcrossing between TSH 858 with Sca 6. The experiment was arranged usingCompletely Randomized Design with four replicates. Cacao seeds wereinoculated by immersing them for 30 minutes in the spore suspension of13 isolates of seed-borne fungi CTB at a density of 10 6 spores/ml. Afterinoculation, the seeds were planted on sterile sand in a plastic box (30 x 30cm). Parameters observed were germination rate, vigor index, KCT-R T 50rate of seedling growth, leaf number, seedling height, root length, rootnumber, and level of pathogenicity. Data were analyzed by ANOVAfollowed with Duncan's Multiple Test. The results showed that the 13species of seed-borne pathogens were in hybrid cacao seeds with varyingpathogenicity. The most pathogenic fungi were Aspergillus flavus,Aspergillus ochraceus, Cladosporium herbanum, Curvularia geniculata,Fusarium oxysporum, Phoma glomerata, and Macrophoma sp. Seed bornepathogenic fungi had the ability to reduce seed germination of 20-40%,vigor index of 30-47%, relative growth rate of 13-45%, and delayedgermination speed (T 50 decreases) from 0.62 to 7.36 days. Seed bornepathogens caused (29-52%) death seed compared to control. All that 13isolates of seed-borne pathogens infected plant tissues such as cotyledons,leaves, stems, and roots of cacao seedlings, but only isolates of Phomaglomerata and Macrophoma sp. which lowered the height of seedlings,leaf number, root number and length. The study indicated that infection ofseed-borne pathogens on cacao seed hybrid can cause seed death.Therefore, seeds should be handled properly.Key words: hybrid seeds, seed borne pathogens, viability, seed vigor,Theobroma caca

Histomorphological changes, sperm quality and testosterone level after administration and cessation of Pegagan (Centella asiatica) extract in rats

The objective of this study was to determine histomorphological change of seminiferous tubules, sperm quality and testosterone level after administration and cessation of pegagan extract. The research used 4x2 factorial complete randomized design, the first factor was dose-administration duration and the second factor was cessation length of pegagan extract. Parameters consisted of degree of spermatogenesis, sperm quality (motility, concentration and abnormality), and testosterone level. The data were processed using analysis of variance and Duncan’s test. Results showed that there was no interaction between dose-administration duration and cessation length. There was no significant difference of dose-administration duration of pegagan on degrees of spermatogenesis, sperm motility and testosterone level. Cessation length did not significantly affect degrees of spermatogenesis, sperm motility, and testosterone level although these results indicated that cessation of pegagan extract for 12 days increased degrees of spermatogenesis and sperm motility, indicated the antifertility potency of pegagan extract. In conclusion, Pegagan extract has potency as anti-fertility agent although the degree of spermatogenesis, sperm quality and testosterone levels were still in the normal range for the reproductive process and not irreversible, and pegagan could be explored as an reversible anti-fertility agent

Successful Intracervical Insemination and Characteristics of Anoa (Bubalus SP.) Parturation Behavior in Captivity

Anoa, which is the endemic animal in Indonesia, its population is unevitably decreasing, therefore anoa included in the list of endangered species by International Union for Conservation of Nature (IUCN). The experiment was aimed to apply the artificial insemination (AI) technique for anoa and to examine their parturition behavior. The experiment involved 2 males and 5 females anoa at Taman Safari Indonesia Bogor. Semen was collected by electroejaculator, then evaluated and diluted in Tris-egg yolk extender. The anesthetized estrous females were inseminated intracervically with extended semen (100x106 sperm/1.0 mL). Females which did not show estrous signs during 2-3 cycles after AI were predicted pregnant, and therefore confirmed by ultrasonography. As a comparison, another predicted-pregnant female after natural mating was scanned. The results showed that intracervical AI resulted pregnancy in one female, and the gestation period of anoa was ranged from 313 d (AI) to 324 d (natural mating). There were three stages of parturition process was observed: Stage 1 was characterized by the abdominal contraction and amniotic membrane rupture, Stage 2 was characterized by the fetal expulsions, and Stage 3 was characterized by the placental expulsions and ingestion of placenta by the dam. The duration of each stage of parturition was 6-8 h (stage 1), 30-60 min (stage 2), and 15-180 min (stage 3). It is concluded that AI technique was applicable to anoa in captivity, and the parturation behavior of anoa was comparable to buffalo and cattle. However, the fetal delivery occurred when the dam was in standing position and the dam ingested the placenta

Cacao (Theobroma Cacao L.) Resistance Evaluation Againts Black Pod Diseaseand Effectiveness of Inoculation Methods

The general objective of this experiment was to standardize method of resistance evaluation of cacao germplasm against Phytophthora palmivora, the pathogen causing black pod disease in cacao. The sepecific objectives were to evaluate (1) effects of inoculum type and pod injury, (2) effects of inoculum type and seedling injury, and (3) effects of genetic background of cacao seedlings on infection of P. palmivora. In this experiment, effectiveness of either zoospora or mycellia was evaluated as inoculation sources for pod, leaf and stem of cacao clone GC 7 and Sca 12. Part of the tested cacao pods, leaves, and stems were injured prior to P. palmivora inoculation while the others were not. Observations were conducted on diameter of the necrosed symptoms on inoculated cacao pods, number of necrosed spot on inoculated leaf, and the length of necrosed symptoms on stem of tested cacao seedlings. Results of the experiment indicated inoculation using mycelia of P. palmivora was more effective than zoospora and injuring the tested cacao pods and seedlings before P. palmivora inoculation can be used to prediction the resistance of tested cacao clones against P. Palmivora infection. Results of resistance prediction using detached pod assay was similar to that of using seedling assay; therefore, seedling assay could be used as an alternattive method of resistance evaluation. Cacao F1 hybrids derived from TSH 858 x Sca 12 showed some resistance against P. palmivora and they might potentially result in high yielding lines