scholarly journals Pewarnaan Kromosom dan Pemanfaatannya dalam Penentuan Tingkat Ploidi Eksplan Hasil Kultur Anter Anthurium

2016 ◽  
Vol 21 (2) ◽  
pp. 113 ◽  
Author(s):  
Budi Winarto
Keyword(s):  
Anther Culture ◽  
Ploidy Level ◽  
Staining Method ◽  
Root Tip ◽  
Root Tips ◽  
Staining Methods ◽  
Ornamental Crops ◽  
High Benefit ◽  
Chromosome Staining

Metode pewarnaan Kromosom yang optimal merupakan prasarat penting dalam penentuan level ploidi tanaman hasil kultur anter, termasuk variasi eksplan hasil kultur anter Anthurium. Aplikasi dan modifikasi metode pewarnaan kromosom pada berbagai eksplan dilakukan di Laboratorium Kultur Jaringan Balai Penelitian Tanaman Hias dari bulan Februari sampai dengan Agustus 2009 untuk mengetahui keragaman dan tingkat ploidi regeneran hasil kultur anter Anthurium. Penelitian bertujuan mendapatkan metode pewarnaan kromosom dan modifikasinya, jenis eksplan dan akar yang sesuai untuk mempelajari tingkat ploidi regeneran hasil kultur anter Anthurium. Bahan yang digunakan ialah kalus, pucuk tunas, dan ujung akar udara. Penelitian terdiri atas tiga kegiatan, yaitu (1) modifikasi metode pewarnaan kromosom, (2) seleksi eksplan yang sesuai untuk pewarnaan kromosom, dan (3) optimasi metode pewarnaan kromosom terseleksi. Hasil penelitian menunjukkan bahwa ujung akar dan akar yang ditumbuhkan pada medium yang mengandung 1% arang aktif merupakan jenis eksplan dan akar yang sesuai untuk mendapatkan hasil pewarnaan kromosom yang baik. Modifikasi metode pewarnaan kromosom dengan pemanasan ujung akar pada 1N HCl : asam asetat glasial 45% (3:1, v/v) selama 10 menit pada suhu 60oC dan perlakuan aseto-orcein selama 15 menit merupakan metode pewarnaan kromosom yang lebih baik dalam menghasilkan obyek kromosom yang mudah dihitung. Penerapan metode pewarnaan kromosom pada kultur anter Anthurium dapat memisahkan tingkat ploidi regeneran. Pada penelitian ini rasio ploidi regeneran kultur anter ialah 33,5% haploid, 62,7% diploid, dan 5,7% triploid. Metode pewarnaan kromosom yang berhasil dikembangkan dalam penelitian ini sangat bermanfaat dalam pengembangan teknologi haploid pada jenis Araceae yang lain.<br /><br />Optimal chromosome staining method is important pre-requisite in determination of plant ploidy level derived from anther culture, involving varied explants regenerated from Anthurium anther culture. Application and modification of chromosome staining methods on different explants were conducted at the Tissue Culture Laboratory of  Indonesian Ornamental Crops Research Institute from February to August 2009 for determination of the ploidy level of regenerants derived from anther culture of Anthurium. The aim of this research was to determine the chromosome staining method and its modifications, type of explant and root suitable to study the ploidy level of explants derived from anther culture of Anthurium. Callus, shoot tips, and root tips were utilized in the experiment. The research was consisted of three experiments, i.e. (1) modification of chromosome staining methods (2) selection of explants suitable for chromosome staining, and (3) improvement of the selected chromosome staining method. Results of the study indicated that root tips and roots cultured on medium containing 1% active carchoal were the most appropriate explants and the root type in obtaining better chromosome staining results. The modification method with root tip boiled in 1N HCl : 45% of acetic acid glacial (3:1, v/v) for 10 minutes in 60ºC and aceto-orcein treatment for 15 minutes gave appropriate chromosome staining results exhibited clearer chromosome pictures and was easy to be counted. The  application of chromosome staining on anther culture of Anthurium was able to distinguish the ploidy level of regenerants. Ploidy ratio of regenerants derived from anther culture was 33.5% of haploid, 62.7% of diploid, and 5.7% of triploid. Chromosome staining method resulted from the study give high benefit in developing haploid technologies on other Araceae plants.<br /><br />

Histochemical Examination of Invertase Activities in the Growing Tip of the Plant Root

2017 ◽  
Vol 10 (1) ◽  
pp. 35-45
Author(s):  
N.F. Lunkova ◽  
N.A. Burmistrova ◽  
M.S. Krasavina
Keyword(s):  
Plant Root ◽  
Invertase Activity ◽  
Root Tip ◽  
Elongation Zone ◽  
Root Tips ◽  
Phloem Unloading ◽  
Meristem Cell ◽  
Transition To Elongation ◽  
Different Tissues

Background:A growing part of the root is one of the most active sinks for sucrose coming from source leaves through the phloem. In the root, sucrose is unloaded from conducting bundles and is distributed among the surrounding cells. To be involved in the metabolism, sucrose should disintegrate into hexoses by means of degrading enzymes.Aims:The aim of this research was to explore the possibility of the involvement of one such enzymes, invertase, in phloem unloading as well as distribution of its activity in the functionally different tissues of the plant root tips.Method:To estimate the enzyme activities in root tissues, we applied two techniques: the histochemical method using nitro blue tetrazolium. The localization of phloem unloading was studied with carboxyfluorescein, a fluorescent marker for symplastic transport.Results:Invertase activity was not detected in the apical part of the meristem. It appeared only between the basal part of this zone and the beginning of the elongation zone. There is the root phloem unloading in that area. Invertase activity increased with increasing the distance from the root tip and reached the highest values in the region of cell transition to elongation and in the elongation zone. The activities of the enzyme varied in different tissues of the same zone and sometimes in the neighboring cells of the same tissue. Biochemical determination of invertase activity was made in the maize root segments coincident to the zones of meristem, cell elongation and differentiation. The results of both methods of determination of invertase activity were in agreement.Conclusion:It was concluded that phloem unloading correlated with invertase activity, possibly because of the activation of invertase by unloaded sucrose. Invertase is one of the factors involved in the processes preparing the cells for their transition to elongation because the concentration of osmotically active hexoses increases after cleavage of sucrose, that stimulates water entry into the cells, which is necessary for elongation growth.

scholarly journals STUDIES ON THE GROWTH HORMONE OF PLANTS

1934 ◽  
Vol 18 (1) ◽  
pp. 23-34 ◽  
Author(s):  
Kenneth V. Thimann
Keyword(s):  
Growth Hormone ◽  
Growth Substance ◽  
Root Tip ◽  
Plant Tissues ◽  
Root Tips ◽  
Direct Extraction ◽  
Simple Method ◽  
Avena Coleoptile ◽  
Method Of Extraction

1. It is shown that when plant tissues are ground with water the growth substance contained therein is inactivated by the oxidizing enzymes. 2. A simple method of extraction is described which enables the quantitative determination of growth substance in such tissues. 3. The amount and distribution of growth substance in the Avena coleoptile is determined by this method, and it is shown that while the substance does not diffuse out from the lower parts of the coleoptile, it is nevertheless present in considerable amounts, the concentration decreasing steadily with the distance from the tip. 4. Growth substance is also present in considerable amounts in Avena roots, and here also its concentration decreases steadily with distance from the tip. 5. The amount of growth substance diffusing out of root tips into dextrose agar, even during long periods of time, is not greater than the amount obtainable by direct extraction. Actual production in the root tip therefore either does not take place at all, or else takes place under quite different conditions from the production in the tip of the coleoptile.

scholarly journals Distribution of Ploidy Level on Adiantumdiaphanum Blume at Various Altitudes in Probolinggo

2019 ◽  
Vol 9 (2) ◽  
pp. 141-149
Author(s):  
Ade Rezi Amelia ◽  
Ahmad Fauzi
Keyword(s):  
Chromosome Number ◽  
Survey Research ◽  
Ploidy Level ◽  
Root Tips ◽  
Altitude Effect ◽  
Number Of Cells

Ferns are indicated could become polyploid organism due to altitude effect. In this research, the distribution of ploidy level on AdiantumdiaphanumBlume based on difference altitude was studied. This study was a survey research conducted in Probolinggo Regency which aims studying the effect of altitude on ploidy level in A. diaphanumBlume. Leces (50 masl), Lumbang (460 masl), and Sukapura (820 masl) were the tree research sampling areas that represent low, medium, and highland location, respectively. The determination of ploidy level was conducted by counting the chromosome number of cells root tips. The results showed that all ferns found in the medium and lowland were diploid organisms, whereas all ferns found in the highlands were triploid organism.

scholarly journals Cytogenetic, chromosome count optimization and automation of Neolamarckia cadamba (Rubiaceae) root tips derived from in vitro mutagenesis

2021 ◽  
Vol 13 (3) ◽  
pp. 10995
Author(s):  
Wei-Seng HO ◽  
Wee-Hiang ENG ◽  
Kwong-Hung LING
Keyword(s):  
Open Source ◽  
Chromosome Count ◽  
Root Tip ◽  
In Vitro Mutagenesis ◽  
Root Tips ◽  
Chromosome Counts ◽  
Laboratory Equipment ◽  
Online Databases ◽  
Chromosome Staining

Chromosome count is the only direct way to determine the number of chromosomes of a species. This study is often considered trivial that seldom described and discussed in detail. Therefore, it is inevitable that the chromosome count protocol should be revised and revisited before it becomes obliterated. In the present study, we encountered challenges in obtaining a clear micrograph for the chromosome count of active mitotic cells of Neolamarckia cadamba (Roxb.) Bosser (Rubiaceae) root tips. Several obstacles were determined through micrograph observation, such as existing unwanted particles in cells, poor chromosome staining and chromosome clumping. To overcome these, root tip types, staining methodologies, squashing methods were among the factors assessed to obtain clear micrographs. The chromosome counts of N. cadamba under optimized procedure showed 2n = 44 chromosomes. We also apply digital technology in chromosome counts, such as online databases and graphic software that are open source and freely accessible to the public. Only basic laboratory equipment and chemicals were used throughout the study, thus making this study economical and applicable in a basic laboratory. The availability of online digital software and databases provide open-source platforms that will ease the efforts in chromosome count.

scholarly journals Studi Tingkat Ploidi pada Lili (Lilium sp.) Hasil Kultur Antera Melalui Penghitungan Jumlah Kloroplas dan Kromosom

Jurnal Agro ◽  
10.15575/864 ◽  
2016 ◽  
Vol 3 (2) ◽  
pp. 34-42
Author(s):  
Dewi Pramanik ◽  
Nisa Istiqomah ◽  
Liberty Chaidir
Keyword(s):  
Anther Culture ◽  
Ploidy Level ◽  
Economic Value ◽  
Ornamental Plant ◽  
F1 Hybrids ◽  
Haploid Plant ◽  
Crop Varieties ◽  
Stomatal Guard Cells ◽  
Ornamental Crops ◽  
Average Standard Error

Lili (Lilium sp.) termasuk famili Liliaceae, merupakan tanaman hias yang memiliki nilai ekonomi tinggi karena permintaan konsumen terus meningkat. Permintaan varietas tanaman yang seragam menuntut pengembangan hibrida F1. Perakitan tanaman hibrida dapat dihasilkan melalui pembentukan tanaman haploid. Salah satu metode untuk memproduksi tanaman haploid adalah dengan kultur antera.  Pengecekan tanaman hasil kultur antera dapat dilakukan dengan penghitungan  jumlah kloroplas dan jumlah kromosom, namun untuk lili hasil belum diperoleh informasi mengenai korelasi antara jumlah kloroplas pada sel penjaga stomata dengan jumlah kromosom, sehingga tujuan penelitian ini adalah untuk mengatahui korelasi antara jumlah kloroplas dengan jumlah kromosom serta mengetahui tingkat ploidi pada regeneran lili hasil kultur antera. Penelitian ini dilaksanakan di Laboratorium Kultur Jaringan Balai Penelitian Tanaman Hias (BALITHI) Cianjur dari Januari-Juni 2016.  Terdapat 5 nomor lili yang diuji tingkat ploidinya.  Setiap nomor terdiri dari 4 ulangan, setiap ulangan ada 4 botol dan  masing-masing botol terdiri dari 3 planlet. Metode analisis yang digunakan yaitu statistik sederhana rata-rata dan standar eror dan dikorelasikan. Hasil penelitian menunjukkan planlet haploid terbanyak ditemukan pada nomor 2015.1.1 kelompok Longiflorum dengan jumlah 26,67% sedangkan planlet haploid yang jumlahnya paling sedikit ditemukan pada nomor 2015.S2.3 kelompok Oriental dengan jumlah 11,11%. Metode kultur antera dapat menghasilkan planlet haploid namun pada Lilium sp. persentase keberhasilannya masih rendah. Tidak diperoleh korelasi antara jumlah kloroplas dan kromosom. Penelitian lebih lanjut terkait tingkat ploidi tanaman lili dan pengujian jumlah kromosom dengan menggunakan flowcytometer perlu dilakukan.  Lily (Lilium sp.), Liliaceae family, is an ornamental plant that has a high economic value as consumer demand continues to rise. The uniformity of crop varieties requires the development of F1 hybrids that can be generated through the formation of haploid plants. A method for producing haploid plant is by anther culture. Evaluation of anther culture can be done by counting the number of chloroplasts and the number of chromosomes in the regenerants.  However, lilies yet obtained information on the correlation between the numbers of chloroplasts in stomatal guard cells with chromosome numbers. Therefore, the purpose of this research aimed to know the correlation between the number of chloroplasts and chromosomes and to determine ploidy level in the regenerants of lily from anther culture. This research was conducted at Tissue Culture Laboratory of Indonesian Ornamental Crops Research Institute (IOCRI), Cianjur from January to June 2016. There were 5 numbers of lilies regenerant from anther culture that evaluate for ploidy level test.  Each number has four replications, each replication contained four bottles, and each bottle has three plantlets. The statistical analysis used statistical descriptive with average, standard error and correlation. The results showed, haploid plantlets were observed in 2015.1.1 plantlets from Longiflorum group with 26,67% while least number of haploid plantlets is found in 2015.S2.3 plantlets from Oriental group (11,11%).  The method of anther culture is able to produce haploid plantlets but the success rate was low in Lilium sp. There were no correlation between the number of chloroplasts and chromosomes.  Further studies related to the ploidy level of lilies from anther culture and the evaluation of chromosomes number by using flow cytometry requires to develop haploid plant of Lily.

scholarly journals AN IMPROVED FIXING AND STAINING PROCEDURE FOR PLOIDY DETERMINATIONS FROM ROOT TIP SQUASHES

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 646e-646
Author(s):  
Henry R. Owen ◽  
A. Raymond Miller
Keyword(s):  
Hydrochloric Acid ◽  
Root Tip ◽  
Staining Procedure ◽  
Chromosome Counting ◽  
Fragaria Ananassa ◽  
Polyploid Species ◽  
Treatment Combination ◽  
Root Tips ◽  
Carbol Fuchsin ◽  
Staining Methods

A comparison of pretreatment, fixing, and staining methods for root tips of Fragaria × ananassa (2n=8x=56), a polyploid species with small chromosomes, was made to facilitate chromosome counting. Three pretreatments (8-hydroxyquinoline, α-bromonaphthalene, and p-dichlorobenzene), three fixatives (Farmer's, Carnoy's, and Newcomer's), and five stains (acetocarmine, lacto-propionic orcein, leucobasic fuchsin, altered carbol fuchsin, and alcoholic hydrochloric-acid carmine) were examined in a factorial design to determine which treatment combination produced the best chromosome preparation. Field propagated runners were grown in sand under greenhouse conditions with supplemental lighting to produce root tips for late morning collection. The treatment combinations of α-bromonaphthalene or 8-hydroxyquinoline, Farmer's fixative, and altered carbol fuchsin, or the combination of α-bromonaphthalene, Farmer's fixative, and alcoholic hydrochloric-acid carmine produced the most intensely-stained and well-defined preparations.

scholarly journals Determination of Cryptosporidium spp. in Van cats by nested PCR

2018 ◽  
Vol 74 (1) ◽  
pp. 6008-2018
Author(s):  
ÖZLEM ORUNÇ KILINÇ ◽  
ALI BILGIN YILMAZ ◽  
YASAR GÖZ ◽  
CUMALI ÖZKAN ◽  
VURAL DENIZHAN
Keyword(s):  
Nested Pcr ◽  
Staining Method ◽  
Health Hazard ◽  
Oral Route ◽  
Adult Males ◽  
Cryptosporidium Spp ◽  
Concentration Method ◽  
Staining Methods ◽  
Stool Samples

The present study was conducted to investigate the presence of Cryptosporidium spp. agents in cats from the Turkish Van Cat Shelter at YüzüncüYıl University by a modified Ziehl-Neelsen staining method and nested PCR. Individual stool samples were obtained from 30 adult females, 30 adult males and 40 kittens – a total of 100 Van cats were analyzed in the study. A simplified formol-ether concentration method was applied to all samples. The samples were then examined microscopically by a modified Ziehl-Neelsen staining method. As a result of the staining, Cryptosporidium spp. oocysts were identified in stool samples of 3 kittens in the microscopic examination. After that, PCR and nested-PCR were conducted with suitable primers. Nested PCR identified 5 kittens (5%) as positive. As a result, it was concluded that nested PCR was a superior diagnostic method for Cryptosporidium diagnosis compared with staining methods and that infected cats could be a health hazard for other cats and individuals, since Cryptosporidium spp. agents infect via the faecal-oral route. Therefore, we believe it is necessary to raise the awareness of people in contact with cats..

Chromosome count improvement and digitalization of Neolamarckia cadamba

2020 ◽  
Author(s):  
Wee-Hiang Eng ◽  
Wei-Seng Ho ◽  
Kwong-Hung Ling
Keyword(s):  
Open Source ◽  
Digital Technology ◽  
Chromosome Count ◽  
Root Tip ◽  
Root Tips ◽  
Chromosome Counts ◽  
Laboratory Equipment ◽  
The Public ◽  
Chromosome Staining ◽  
Graphic Software

Abstract Background: Chromosome count is the only direct way to determine the number of chromosome of a species. This study is often considered trivial that seldom described and discussed in detail. Therefore, it is evitable that chromosome count protocol should be revised and revisited before it becomes obliterated. Our chromosome counts in N. cadamba root tips have encountered challenges that prevent us from obtaining clear micrograph with the correct chromosome number. Several obstacles were determined through micrograph observation such as existing unwanted particles in cells, poor chromosome staining and chromosome crumping. Results: The chromosome counts of Neolamarckia cadamba under optimized procedure yielded 44 chromosomes per nucleus. To overcome these, root tip types, stain types, squashing methods were among factors assessed to obtain clear micrograph. We also apply digital technology in term of online database and graphic software that are open source and freely accessible to the public. Conclusions: Throughout the study, we used a few basic laboratory equipment and chemicals, thus making this study economical and applicable in a basic laboratory. The availability of online digital software and databases provide open-source platforms that will ease the efforts in chromosome count.

Laboratory Techniques for Determining Ploidy Levels

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 555c-555
Author(s):  
Christopher S. Cramer
Keyword(s):  
Ploidy Level ◽  
Root Tip ◽  
Ploidy Levels ◽  
Ploidy Determination ◽  
Course Of Action ◽  
Stomata Size ◽  
Pollen Grain Size ◽  
Ploidy Chimeras ◽  
Polyploid Crops

The determination of ploidy levels is essential for breeding asexually propagated and polyploid crops that may have ploidy chimeras in their tissues or have several polyploid series. This presentation will discuss a laboratory for teaching students how to determine ploidy levels in different plant tissues using different techniques. The different methods for ploidy determination include root tip squashes, pollen mother cell squashes, pollen grain size and germinal pore counts, stomata size and density determination, and gross morphology. After completing these laboratory experiments, students will be able 1) to initiate a study of ploidy determination, 2) to determine the correct course of action in determining ploidy level, and 3) to recognize the various steps required to determine ploidy level in plants.

scholarly journals Tendrils as an Alternate Tissue Source for Chromosome Visualization

1994 ◽  
Vol 119 (4) ◽  
pp. 850-852 ◽  
Author(s):  
R.C. Yadav ◽  
Rebecca Grumet
Keyword(s):  
Pisum Sativum ◽  
Vitis Vinifera ◽  
Ploidy Level ◽  
Cucumis Melo ◽  
Root Tip ◽  
Root Tips ◽  
Tissue Source ◽  
Preparation Techniques

Tendrils were used as an alternate tissue source to determine the ploidy level of transgenic melon (Cucumis melo L.) plants. Young tendril tips were prepared for chromosome visualization using standard root tip preparation techniques. Well-spread chromosome preparations were obtained. Tendrils were found to be as amenable to chromosome visualization as root tips. The transformed melon plants, representing three transgenic melon lines, were tetraploid. The tendril technique also was applied successfully to several other tendril-producing species: cucumber (C. sativus L.), pea (Pisum sativum L.), and grape (Vitis vinifera L.). This technique can be very useful for chromosome visualization in tendril-producing species, and can be especially valuable when root tips or anthers cannot be obtained easily.

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