Chloroplast DNA Copy Number May Link to Sex Determination in Leucadendron (Proteaceae)

Leucadendron (Proteaceae) is a South African genus, the flowers of which have become a popular item in the Australian cut-flower industry. All species are dioecious. In general the female flowers are the more desirable as cut flowers. The availability of a molecular marker linked to sex determination is therefore needed both to maximize the efficiency of breeding programs and to supply markets with flowers from the preferred sex. The polymerase chain reaction-based method of suppression subtractive hybridization (SSH) combined with mirror orientation selection (MOS) were applied in an attempt to identify genome differences between male and female plants of Leucadendron discolor. Screening of 416 clones from a male-subtracted genomic DNA library and 282 clones from a female-subtracted library identified 13 candidates for male-specific genomic fragments. Sequence analyses of the 13 candidate DNA fragments showed that they were fragments of the chloroplast DNA, raising the possibility that chloroplast DNA copy number is linked to sex determination in Leucadendron. Key words: Leucadendron, sex determination, suppression subtractive hybridization (SSH

PCR-RFLP and Sequencing of TrnS/trnfM Fragment of Enhalus Acoriodes From Sanur Coastal Waters, Bali, Indonesia: a Preliminary Study

This study aimed to evaluate the resolution of PCR-RFLP and sequencing of trnS/trnfM in detecting genetic diversity of seagrass Enhalus acoroides at Sanur Beach, Sindhu Beach and Semawang Beach. This research used six samples E. acoroides from each location. DNA extraction method followed Doyle and Doyle. PCR amplifications were done using primers P672/P673, P674/P675 and ITS4/ITS5 followed by digestion using restriction enzymes EcoRI, PstI, HindIII, BamHI, Rsa, Mva and HinfI. Sequencing analysis used PCR products of primers trnS/trnfM. Testing of DNA sequences of E. acoroides were conducted using BLAST (Basic Local Alignment Search Tool). DNA sequences were further analyzed using MEGA 5.2 software (Molecular Evolutionary Genetic Analysis) to evaluate variations of DNA. The sequence alignments were done using ClustalW software to determine the homology between the DNA sequences. The results showed that 18 samples of E. acoroides from Sanur, Sindhu and Semawang Beach have no polymorphism based on restriction enzyme analyses. Furthermore, sequencing of trnS/trnfM region of 18 samples E. acoroides showed that the sequences were identical

Study of the natriuretic peptide hormone system in plants

In this study, both physiological and cellular effects are elicited by natriuretic peptides (NPs), a novel type of plant hormone. It was found that rat ANP (rANP) influenced stomatal opening movement in Tradescantia sp., where a significant increase in stomatal opening was observed in the presence of 1 µM rANP. Furthermore, this effect is mediated by cGMP, a (putative) second messenger of NPs. Two inhibitors of guanylyl cyclase, LY 83583 and methylene blue, inhibited rANP-induced stomatal opening. In contrast, stomatal opening is induced in a concentration dependent manner by the cell permeant cGMP analogue 8-Br-cGMP. In addition it was found, that like in animals, the secondary structure of rANP is essential for rANP responses. Linearised rANP is biologically inactive. Since ANP elicit plant responses, an attempt was made to isolate NP analogues from plants. A protocol for partially purifying NP from plants was developed. It was found that two fractions eluted from an immunoaffinity chromatography column (0.5 M KCI eluted fraction and 0.75 M KCI eluted fraction) were biologically active. The level of cGMP in response to NPs was also tested. It is suggested that the receptor of NP is specific since only 0.75 M KCI eluted fractions increased cGMP levels in Zea mays root stele tissue. rANP did not elicit an effect on cGMP levels in this tissue and LY 83583 did not affect this response. It is therefore argued that a plant specific biologically active NP system is present in the stele and it is predicted that NPs modulate solute movement in this tissue. NPs also influence K+, Na+ and H+ fluxes in Zea mays root stele. Increase in both K+ and Na+ uptake were observed after 30 min., while H+ flux shifted immediately toward influx in the presence of both 0.5 and 0.75 KCI eluted fractions. Finally, a model is proposed for the effect of NPs on solute movement and its signalling system in plants

RAPD Primers Selection for Genetic Variation Analysis of Banana Plant (Musa spp.)

Identification of genetic diversity using molecular markers is important for basic information for plant conservation. Banana is a fruit bearing plants that important for food sources in human life. This study aimed to determine the optimum conditions of  PCR-RAPD reaction and RAPD primers that suitable to amplify DNA fragmens. DNA isolation was done using modified of CTAB and chloroform isoamil alcohol. The samples used was young leaves of nine banana cultivar plants. Optimation was done using variety of DNA and MgCl2 concentration. Eight primers produced by Operon Primer Technology were tested. The DNA genomic concentration obtained was in the range of 23,3 ng/µl – 70 ng/µl. The optimum conditions of PCR-RAPD of banana plants that produce clear band were 50 ng/µl DNA template, 3 mM MgCl2 with the number of thermal cycles was 40 x. There were six RAPD primers that successfully amplified DNA : OPA 02, OPA 04, OPB 12, OPD 20, OPH 01, and OPH 03. The primer OPA-04 had the lowest resolving power value (4,4) , while  OPH 01 had the highest (11,3) resolving power

Genetic Diversity and Population Structure of Bullet Tuna (Auxis rochei) from Bali and Its Adjacent Waters

Bullet tuna (Auxis rochei) dominates the neritic tuna catch, especially from the purse seine fleet within the western and southern Indonesian waters. However, high catches can lead to stock depletion and lower genetic diversity due to possible inbreeding. Therefore, population genetic information is important in monitoring the sustainability of fish stocks and proposing an appropriate species-specific conservation strategy. This study aimed to analyze the genetic diversity, population structure, and kinship relationship of bullet tuna in Bali and its adjacent waters. Sampling was carried out in September 2020 at landing sites/ports representing the north, east, south, and west region, whereas at least 30 samples were acquired at each location. The result showed that the DNA concentration obtained could produce DNA bands with allele length ranged from 94-260 bp. Observed heterozygosity (Ho) was around 0.440-0.627. While the expected heterozygosity (He) was between 0.932-0.945. The genetic variation among population, within-population, and individuals was 0.36%, 41.04%, and 58.60%, respectively. The results of the analysis of genetic diversity between individuals in the population showed very high genetic diversity. The population structure of the bullet tuna landed in West Bali, East Bali, South Bali and North Bali is the same population stock. The kinship relationship indicates that the four populations are closely related genetically

VALUASI NILAI EKONOMI WISATA PANTAI SAWARNA DENGAN MENGGUNAKAN TRAVEL COST METHOD

Pantai Sawarna di Kabupaten Lebak memiliki keindahan alam yang dapat digunakan sebagai daya tarik objek wisata bagi setiap pengunjung. Keindahan alam pada kenyataannya tidak terhitung menjadi nilai ekonomi atau tidak memiliki nilai pasar, sehingga pentingnya dilakukan valuasi ekonomi untuk memperkecil eksternalitas yang dapat menyebabkan kegagalan pasar karena suatu barang atau jasa yang tidak memiliki nilai pasar, sehingga dalam kajian ini bertujuan untuk menentukan nilai ekonomi Pantai Sawarna dari sumberdaya alam yang tidak memiliki nilai Pasar. Metode valuasi ekonomi yang digunakan adalah metode biaya perjalanan dengan pendekatan individu (Individual Travel Cost Method). ITCM dipilih sebagai pendekatan yang lebih teliti dalam menilai kondisi sosial ekonomi pengunjung sehingga mengetahui nilai ekonomi dan nilai Willingness To Pay dari biaya perjalanan pengunjung. Willingness To Pay (WTP) dijadikan untuk mencari nilai ekonomi dan surplus konsumen area Pantai Sawarna. Hasil dari valuasi ekonomi dengan pendekatan ITCM yang digunakan di area Pantai Sawarna tersebut didapat nilai rata-rata WTP sebesar Rp. 93.672,-/pengunjung, dan nilai ekonomi Pantai Sawarna adalah Rp. 6.722.558.424,- / tahun. Nilai surplus konsumen adalah Rp. 5.164.066.252,-

The Genetic Relationships of Grevillea Hybrids Determined by RAPD Marker

Grevillea (Proteaceae) is native to Australia and is known as garden plant. The flowers have high diversity colors, which range over orange, yellow, pink, red and green. It is a large genus, consisted of 357 species. This genus is economically important and there are hundreds of Grevillea hybrids with beautiful color and shape of flower.  Information of the genetic relationships of Grevillea hybrids is not available. Ten Grevillea hybrids were used in this study to evaluate the genetic relationships between hybrids and with their parents. PCR-RAPD technique was employed in this study with seven RAPD primers. The dendrogram of Grevillea hybrids was performed using Neighbor-Joining analysis based on genetic distance. The analysis revealed that ‘Moonlight’, ‘Caloundra Gem’, and Little Pink Willie’ were clustered with Grevillea banksii. The three hybrids were hybrid of G. banksii and other Grevillea species.  ‘Superb’ was closely related to ‘Robyn Gordon’ as they are hybrid of G. banksii and G. bipinnatifida

PENGEMBANGAN PENGELOLAAN SISTEM PERSAMPAHAN AREA WISATA GRAMA TIRTA JATILUHUR MENGGUNAKAN PRINSIP CONTINGENT VALUATION METHOD

Area Wisata Grama Tirta Jatiluhur (GTJ) merupakan suatu destinasi wisata yang hampir seluruh fasilitas yang disediakan mengandalkan keindahan alam sekitar Waduk dan menjadikannya daya tarik utama bagi setiap pengunjung, yang pada kondisinya tidak terhitung menjadi nilai ekonomi area wisata GTJ karena tidak memiliki nilai pasar. Valuasi ekonomi dibutuhkan untuk memperkecil dampak dari kegagalan pasar akibat tidak memilikinya nilai guna sehingga menjadikan peluang terjadinya eksploitasi pada lingkungan  menyebabkan pencemaran lingkungan terjadi di area wisata GTJ. Pencemaran lingkungan di area wisata GTJ disebabkan oleh sampah yang dibuang tidak pada tempatnya karena kurang baiknya sistem pewadahan dan pengumpulan sampah. Pendekatan yang digunakan dalam valuasi ekonomi adalah Contingent Valuation Method (CVM) untuk menngukur nilai keberadaan dari sumber daya alam yang di eksploitasi dan menyebabkan pencemaran. Prinsip CVM yang digunakan adalah pasar hipotesa, penawaran pengembangan sistem pewadahan dan persampahan  Willingness to Pay (WTP) pengunjung. Penggunaan prinsip tersebut memberikan solusi untuk pengembangan pengelolaan sistem persampahan khususnya sistem pewadahan dan pengumpulan sampah. WTP yang didapat dijadikan dasar biaya sistem pewadahan dan pengumpulan sampah. WTP total yang didapat adalah Rp.988.330.500 dengan WTP rata-rata pengunjung adalah Rp.4.500

Optimasi formula krim body scrub ekstrak lamun (Enhalus acoroides) terenkapsulasi

Body scrub cream is used to clean and nourish the skin. Several illegal creams were reported to contain harmful chemicals. Some plant extracts can replace chemical compounds because they can lighten the skin. Enhalus acoroides is one of the seagrass species that is widely distributed in Indonesia. Leaf extract of E. acoroides was found to contain phenolic, tannin, and flavonoid. Due to the bioactive compounds in the E. acoroides extract can be added to cosmetic products such as body scrub cream. This study aimed to analyze the effect of the addition of encapsulated E. acoroides extract on the characteristics of the body scrub cream and determined the concentration of the encapsulated E. acoroides extract, which produced the best body scrub cream. The extraction was done using chloroform: ethanol (9:1) and encapsulation using maltodextrin and tween. The body scrub cream was made in two phases, the oil phase and the water phase. The concentrations of the encapsulated E. acoroides extract tested were 0%, 0.5%, 1%, 1.5%, 2%, 2.5%. The characteristics evaluated were homogeneity, pH, viscosity, spreadability, stickiness, IC50, and organoleptic test for color and scent. The results showed that the body scrubs were homogenous. The pH values were 5.43-6.66, and viscosity was 34400-46400cp which are in the range of SNI for cosmetic products. The spreadability values were 4.03-5.5cm, and stickiness was 4.8-6.91sec. The antioxidant capacity (IC50) ranged from 147.38 to 157.64 ppm, and the overall acceptance scores were 5 to 6.80. Based on the effectivity index and statistical analyses of physical-chemical characteristics, the best concentration of encapsulated E. acoroides extract was 2%.  These findings indicate the potential of encapsulated E. acoroides extracts to improve skin health

Pollen Viability and Microspore Culture in Three Broccoli Cultivars (Brassica oleracea L. var. italica Plenck)

Broccoli is a high value vegetable crop in Indonesia, however production is low due to limited number of suitable cultivars, so, breeding hybrid broccoli for warm climate is important.  The first step in hybridization is providing homozygote parent plants which can be done efficiently via microspore culture.  The objectives of this study were to determine 1) bud size that produce uninucleate microspore stage appropriate for culture; 2) pollen viability, 3) microspore development, in three broccoli cultivars (‘BL 10001’, ‘Royal Green’ and ‘Green Magic’). Various bud size (1 – 5 cm) was squashed and observed microscopically to determine bud size containing uninucleate microspore.  Pollen viability was determined by IKI staining and pollen germination method.  Chromosome number was counted on root tips using squash method with aceto-orcein stain. Various heat treatment schemes were conducted to induce microspreo development.  Result showed uninucleate microspore derived from 2-3 mm and 3-4 mm bud length of ‘BL-10001’ and ‘Royal Green’ was responsive for microspore development in culture.  Pollen viability varied among cultivars, 78-87% on IKI method and 15-16% on germination test. Microspore culture showed different embryogenesis response; pollen-like structure was produced by ‘BL 10001’