Hybrid Triploid Induced by Megaspore Chromosome Doubling in Jujube (Ziziphus jujuba Mill.) ‘Maya’ and Its Characteristics

Polyploid breeding is an important strategy for tree improvement because polyploid individuals typically show superior traits, such as improved growth, stress resistance, and superior fruit quality. Artificial induction of chromosome doubling of female gametes is an effective approach to obtain triploid progeny. However, no triploid fruit tree cultivars have been developed using this approach. The objective of this study was to explore the utility of chromosome doubling in female gametes of ‘Maya’ jujube to produce triploid individuals. The temporal relationship between flower bud morphology and the megaspore meiotic stage was studied to guide the optimal timing of colchicine treatment. Colchicine solution was applied to bearing shoots of mature ‘Maya’ jujube trees in a field experiment using two treatment methods (improved cotton leaching and injection method) and three concentrations (0.3%, 0.4%, and 0.5%). The water transport rate of ‘Maya’ jujube shoots was studied using dye solution to judge the effectiveness and timing of the colchicine treatment methods. Two triploids were identified among the progenies from the colchicine-treated shoots. The highest efficiency of triploid production was 3.3% when flower buds of diameter 1.76–2.12 mm were treated with 0.3% colchicine solution for 4 h using an improved cotton leaching method. The ground diameter, plant thorn length, leaf width, leaf area, stomatal length, stomatal width, chlorophyll content, and photosynthetic parameters of one triploid individual were significantly higher than those of diploids of identical parentage at 18 months old. Thus, induction of 2n megaspores is an effective approach to generate triploid jujube. These results are expected to promote and accelerate triploid breeding in fruit trees.

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Colchicine-induced Amphidiploids of Rose Interspecific Hybrids

HortScience ◽

10.21273/hortsci.30.4.778a ◽

1995 ◽

Vol 30 (4) ◽

pp. 778A-778

Author(s):

Yan Ma ◽

David H. Byrne ◽

Jing Chen

Keyword(s):

Interspecific Hybrids ◽

Chromosome Doubling ◽

Diploid Species ◽

Colchicine Treatment ◽

Transfer Resistance ◽

Breeding Research ◽

Rose Breeding ◽

Colchicine Solution ◽

Ploidy Chimeras

An objective of our rose breeding research is to transfer resistance to blackspot and other diseases from wild diploid species to modern rose cultivars. Interspecific hybrids among blackspot-resistant diploid species were chosen for chromosome doubling to produce fertile amphidiploids that could be hybridized to the tetraploid commercial germplasm. Five such F1 interspecific hybrids were treated with colchicine. The study included two different application procedures (shake in colchicine solution or colchicine in media), four colchicine concentrations (0.05%, 0.1%, 0.15%, and 0.20%), and five treatment periods (1, 3, 5, 8, and 10 days). After colchicine treatment, all the materials were cultured in vitro. One thousand-thirty-seven surviving explants were selected for typical “gigas” characteristics of doubled diploids. Chromosome counts on shoot tips of these selected genotypes confirmed 15 amphidiploids. The best colchicine treatment varied among the interspecific hybrids. Higher colchicine concentrations or duration reduced growth rating, rooting, and percent survival. The recognition of amphiploids and ploidy chimeras from young seedlings will also be discussed.

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Megaspore Chromosome Doubling in Eucalyptus urophylla S.T. Blake Induced by Colchicine Treatment to Produce Triploids

Forests ◽

10.3390/f9110728 ◽

2018 ◽

Vol 9 (11) ◽

pp. 728 ◽

Cited By ~ 3

Author(s):

Jun Yang ◽

Jianzhong Wang ◽

Zhao Liu ◽

Tao Xiong ◽

Jun Lan ◽

...

Keyword(s):

Tree Species ◽

Chromosome Doubling ◽

Treatment Plant ◽

Solution Treatment ◽

Colchicine Treatment ◽

Treatment Technique ◽

Chromosome Counting ◽

Triploid Breeding ◽

Optimal Period ◽

Colchicine Solution

Triploids generally provide an advantage in vegetative growth in forest trees. However, the technique of triploid breeding is still an open field in the Eucalyptus tree species. This study aims to explore the colchicine treatment technique for megaspore chromosome doubling to establish triploids in this tree species. Cytological observation on microsporogenesis and megasporogenesis was carried out to guide megaspore chromosome doubling induced by colchicine treatment. Ploidy level in progenies was detected by flow cytometry and somatic chromosome counting. A relationship between microsporogenesis and megasporogenesis was established to guide the colchicine treatment. Seven triploids were obtained in the progenies, and the highest efficiency of triploid production was 6.25% when the flower buds underwent a 0.25% colchicine solution treatment for 6 h using an aspiration method seven days after the first observation of leptotene during microsporogenesis on the floral branch. Cytological analysis showed that the megasporocyte from leptotene to diakinesis may be the optimal period for megaspore chromosome doubling by colchicine treatment. Plant height, ground diameter, leaf area, and the photosynthetic parameter of triploid eucalypt were significantly higher than those of the diploid plant at 6 months old. Hybridization with 2n megaspores induced by colchicine treatment is an effective way for Eucalyptus triploid breeding. These results should accelerate the development of advanced germplasms in this tree species.

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Putative diploid ancestors of 80-chromosome Glycine tabacina

Genome ◽

10.1139/g92-023 ◽

1992 ◽

Vol 35 (1) ◽

pp. 140-146 ◽

Cited By ~ 12

Author(s):

R. J. Singh ◽

K. P. Kollipara ◽

F. Ahmad ◽

T. Hymowitz

Keyword(s):

Adventitious Roots ◽

Chromosome Doubling ◽

Colchicine Treatment ◽

Meiotic Pairing ◽

B Genome ◽

Specific Hybridization ◽

A Genome ◽

Genome Homology ◽

Glycine Tabacina ◽

Natural Mutation

The objective of this study was to discover the diploid progenitors of 80-chromosome Glycine tabacina with adventitious roots (WAR) and no adventitious roots (NAR). Three synthetic amphiploids were obtained by somatic chromosome doubling. These were (i) (G. latifolia, 2n = 40, genome B1B1,) × (G. microphylla, 2n = 40, genome BB) = F1(2n = 40, genome BB1) – 0.1% colchicine treatment (CT) – 2n = 80, genome BBB1B1; (ii) (G. canescens, 2n = 40, genome AA) × G. microphylla, 2n = 40, genome BB) = F1 (2n = 40, genome AB) – (CT) – 2n = 80, genome AABB; (iii) (G. latifolia, 2n = 40, B1B1) × G. canescens, 2n = 40, AA) = F1 (2n = 40, genome AB1) – (CT) – 2n = 80, genome AAB1B1. The segmental allotetraploid BBB1B1 was morphologically similar to the 80-chromosome G. tabacina (WAR), but meiotic pairing data in F1 hybrids did not support the complete genomic affinity. Despite normal diploid-like meiosis in allotetraploids AABB and AAB1B1, AABB was completely fertile, while pod set in AAB1B1 was very sparse. Morphologically, allotetraploid AABB was indistinguishable from the 80-chromosome G. tabacina (NAR) but in their F1 hybrids, the range of univalents at metaphase I was wide (4–44). The allotetraploid AAB1B1 did not morphologically resemble the 80-chromosome G. tabacina (NAR). However, the F1 hybrid of AABB × AAB1B1 showed normal meiosis with an average chromosome association (range) of 1.7 I (0–4) + 39.2 II (38–40). Based on this information, we cannot correctly deduce the diploid progenitor species of the 80-chromosome G. tabacina (NAR). The lack of exact genome homology may be attributed to the geographical isolation, natural mutation, and growing environmental conditions since the inception of 80-chromosome G. tabacina. Thus, it is logical to suggest that the 80-chromosome G. tabacina (NAR) is a complex, probably synthesized from A genome (G. canescens, G. clandestina, G. argyrea, G. tomentella D4 isozyme group) and B genome (G. latifolia, G. microphylla, G. tabacina) species, and the 80-chromosome G. tabacina (WAR) complex was evolved through segmental allopolyploidy from the B genome species.Key words: Glycine spp., allopolyploidy, colchicine, genome, intra- and inter-specific hybridization, polyploid complex.

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Transcriptome analysis reveals plant response to colchicine treatment during on chromosome doubling

Scientific Reports ◽

10.1038/s41598-017-08391-2 ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 5

Author(s):

Kai Zhou ◽

Paige Fleet ◽

Eviatar Nevo ◽

Xinquan Zhang ◽

Genlou Sun

Keyword(s):

Transcriptome Analysis ◽

Chromosome Doubling ◽

Colchicine Treatment ◽

Plant Response

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Colchicine-induced chromosome doubling in Pennisetum interspecific hybrids and its effect on plant morphology

Indian Journal of Genetics and Plant Breeding (The) ◽

10.31742/ijgpb.80.1.7 ◽

2020 ◽

Vol 80 (01) ◽

Author(s):

Arshpreet Kaur ◽

Rahul Kapoor ◽

Yogesh Vikal ◽

Anu Kalia

Keyword(s):

Interspecific Hybrids ◽

Biomass Yield ◽

Chromosome Doubling ◽

Plant Morphology ◽

Colchicine Treatment ◽

Stem Cuttings ◽

Chromosome Counting ◽

Stomatal Frequency ◽

Preliminary Screening ◽

Stomata Size

We report the production of hexaploid plants of interspecific hybrids of Pennisetum, with the ultimate aim to improve the biomass yield, drought tolerance and multicut behaviour of this genus. Chromosome doubling was achieved with the application of colchicine at three different concentrations (0.05, 0.1 and 0.2%) for two time durations (12 and 24 hours). The root slips and stem cuttings of interspecific hybrids were used for treatment and the root slips were found to be more efficient. The preliminary screening to select the putative hexaploid plant was done based on stomatal frequency and morphology. Plants containing significantly lower stomatal frequency and larger stomata size were selected for further analysis by chromosome counting. This experiment confirmed that 0.1% concentration of colchicine treatment to root slips for 24 hours was more effective to induce the amphiploids in Pennisetum.

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Production and Identification of a Tetraploid Germplasm of Edible Lilium davidii var. unicolor Salisb via Chromosome Doubling

HortScience ◽

10.21273/hortsci12004-17 ◽

2017 ◽

Vol 52 (7) ◽

pp. 946-951 ◽

Cited By ~ 3

Author(s):

Yayan Feng ◽

Leifeng Xu ◽

Panpan Yang ◽

Hua Xu ◽

Yuwei Cao ◽

...

Keyword(s):

Narrow Range ◽

Chromosome Doubling ◽

Colchicine Treatment ◽

Guard Cells ◽

Growth Increment ◽

Low Fertility ◽

Genetic Characteristic ◽

New Germplasm ◽

Annual Growth Increment ◽

Lilium Davidii

Lilium davidii var. unicolor Salisb is a cultivar of Lilium (Liliaceae) with important edible and ornamental characteristic. The application and production of Lilium davidii var. unicolor Salisb were still facing large problems because of its several disadvantages such as narrow range of adaptability, small annual growth increment, and low fertility. To achieve broader environmental adaptability and obtain a more nutritious germplasm, we used colchicine and oryzalin to induce chromosome doubling via the soaking method. Tissue culture bulbs were treated with colchicine at 0.03%, 0.05%, or 0.08% for 32, 40, or 48 hours or with oryzalin at 0.002%, 0.005%, 0.008%, or 0.01% for 3, 6, 9, 12, or 24 hours before being transferred to a differentiating medium. The results showed that colchicine treatment resulted in the highest induction rate when applied at 0.05% for 48 hours, whereas oryzalin treatment produced fewer tetraploid plants. The chromosome number of induced plants with small stoma density and longer guard cells is twice than that of the diploid. The plants were identified as tetraploid. In this study, a new germplasm of Lilium davidii var. unicolor Salisb was innovative and showed novel genetic characteristic.

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Chromosome Doubling in Limonium bellidifolium (Gouan) Dumort. by Colchicine Treatment of Seeds

The Horticulture Journal ◽

10.2503/hortj.mi-117 ◽

2016 ◽

Vol 85 (4) ◽

pp. 366-371 ◽

Cited By ~ 5

Author(s):

Shiro Mori ◽

Takuya Yamane ◽

Masaki Yahata ◽

Koichi Shinoda ◽

Naho Murata

Keyword(s):

Chromosome Doubling ◽

Colchicine Treatment

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Nucleotypic influences on chromosome-specific chiasma variation in Crepis capillaris. I. Responses to early colchicine treatment and chromosome doubling

Genome ◽

10.1139/g96-135 ◽

1996 ◽

Vol 39 (6) ◽

pp. 1078-1085 ◽

Cited By ~ 1

Author(s):

M. T. Abberton ◽

R. S. Callow

Keyword(s):

Key Words ◽

Chiasma Frequency ◽

Chromosome Doubling ◽

Colchicine Treatment ◽

Seedling Stage ◽

Published Data ◽

Crepis Capillaris ◽

The Mean ◽

Diploid Level ◽

Multiplicative Effects

Additive and multiplicative effects of colchicine treatment at the seedling stage and of chromosome doubling on chromosome-specific chiasma frequency at metaphase I have been measured in comparisons between C0 and untreated diploids and between C0 autotetraploids and C0 diploids. Early colchicine treatment increases the frequency of chromosome C univalents to 1.8% but has no similar effect on chromosomes A and D. Colchicine treatment has little net effect on mean chiasma frequency, deducting an average of 0.204 chiasmata per set but otherwise multiplying the mean by a factor of 1.182. These additive and multiplicative effects represent averages of six phenotypes. Chromosome doubling in a tetraploid–diploid chimaera subtracts an average of 0.265 chiasmata per set but otherwise doubles the numbers of chiasmata at the diploid level (× 2.134). Comparison of six diploids and tetraploids reveals modest average additive (+ 1.103) and multiplicative effects (× 1.190). The implications of these findings are discussed in the light of new analyses of previously published data. Key words : chiasmata, Crepis, colchicine, meiosis, polyploidy.

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PENGARUH KOLKHISIN TERHADAP PENAMPILAN LADA P p er nigrum L.) MUTAN DAN ANALISIS PLOIDI

Jurnal Penelitian Tanaman Industri ◽

10.21082/littri.v21n3.2015.125-130 ◽

2016 ◽

Vol 21 (3) ◽

pp. 125

Author(s):

NATALINI NOVA KRISTINA ◽

SITTI FATIMAH SYAHID

Keyword(s):

Genetic Variability ◽

Growth Parameters ◽

Colchicine Treatment ◽

Piper Nigrum ◽

Green House ◽

Ploidy Analysis ◽

Plant Phenotype ◽

Medicinal Crops ◽

Colchicine Solution ◽

Research Institute

ABSTRAKKeragaman genetik plasma nutfah lada (Piper nigrum) di Indonesia rendah sehingga perlu dilakukan peningkatan keragaman. Penelitian dilakukan di Rumah Kaca Balittro Bogor mulai Januari 2012 sampai Juni 2013. Tujuan penelitian adalah untuk meningkatkan keragaman genetik lada varietas Petaling 1 menggunakan mutagen kimia kolkhisin. Biji lada direndam dalam larutan kolkhisin konsentrasi 0; 0,01; 0,03; dan 0,05% selama 4 jam dan disemai pada bak pasir. Masing-masing perlakuan diulang 3 kali dan setiap ulangan terdiri atas 300 biji. Pengamatan dilakukan terhadap persentase perkecambahan dan fenotipe tanaman, persentase tumbuh, tinggi tanaman, serta jumlah ruas dan daun pada umur dua bulan. Selanjutnya, sebanyak 20 individu dari total benih yang tumbuh dipilih berdasarkan rata rata penggabungan dari tanaman terpendek dan tertinggi. Individu terpilih diamati tinggi tanaman serta jumlah ruas dan daun pada umur empat bulan. Untuk melihat ragam genetik dilakukan analisis kandungan DNA dengan flowcytometry. Hasil penelitian menunjukkan bahwa perlakuan kolkhisin 0,01 dan 0,05% menghasilkan persentase perkecambahan benih di persemaian lebih tinggi. Pada lada mutan vegetatif generasi ke-0, perubahan morfologi terindikasi pada konsentrasi 0,03%. Pada generasi mutan hasil perbanyakan/turunan vegetatif pertama perubahan morfologi pada tanaman terjadi pada perlakuan 0,05%. Namun, tidak ada perbedaan yang nyata pada tingkat ploidi lada pada semua perlakuan termasuk kontrol.Kata kunci: Piper nigrum L., ragam genetik, mutan, kolkhisin, fenotip ABSTRACTEffect of Colchicine on the Phenothypic Performance of Pepper (Piper nigrum L.) Mutant and Ploidy analysisGenetic variability of pepper (Piper nigrum) in Indonesia was low, so it was needed to increase its variability. Research was conducted at the green house of Indonesian Spices and Medicinal Crops Research Institute, Bogor from January 2012 to June 2013. The aim of the research was to increase the genetic variability of pepper (Petaling 1) using chemical mutagen colchicine. Seeds of pepper were soaked in colchicine solution with several concentration (0; 0,01; 0,03; and 0,05%) for four hours, and then germinated on sand media. Every treatment consisted of 300 seeds and replicated three times. The parameter observed were germination percentage, plant phenotype, growth percentage, plant hight, number of node and leaves two months after planting. Further, from total seedling growth, 20 individual were selected based on average combined from highest and shortest plant. The selected individual observed their plant height, number of node and leaves on four months. Flowcytometri analysis from the selected seedling was conducted to find interplant genetic variabilities. The result showed that application of colchicin 0,01 and 0,05% performed the fast germination on the nursery compared with control, but no significant differencet on the growth parameters. In the mutant generation 0, the changes on morphology showed on 0,03% and at the first vegetative generation, the changes were indicated in plants from 0,05% of colchicine treatment. Flowcytometri analysis showed no significant differences on ploidi level of all treatments including control.Keywords: Piper nigrum, genetic variability, mutant, cholchicin, phenotype

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Induksi PLB Anggrek Vanda sumatrana Schltr. Liar Pada Media MS dengan Penambahan BAP dan NAA serta Ploidisasi dengan Kolkisin

JURNAL BIOLOGI UNAND ◽

10.25077/jbioua.4.4.208-215.2015 ◽

2015 ◽

Vol 4 (4) ◽

pp. 208

Author(s):

Hanifah - Aini ◽

Mansyurdin - Mansyurdin ◽

Suwirmen - Suwirmen

Keyword(s):

Plant Physiology ◽

Tissue Culture ◽

Acetic Acid ◽

Survival Rate ◽

Cell Biology ◽

Colchicine Treatment ◽

Ms Medium ◽

Colchicine Solution ◽

Department Faculty

The study about PLB induction of wild Vanda sumatrana Schltr. on MS media suplement with BAP and NAA and ploidisation by colchicine treatment was conducted from December 2014 until November 2015 at the Laboratory of Genetics and Cell Biology and Laboratory of Plant Physiology and Tissue Culture, Biology department, Faculty of Mathematic and Natural Science, Andalas University, Padang. The study aimed to 1) knowing the best concentration of 6-Benzyl amino purin (BAP) and α-Naphtalene acetic acid (NAA) for Protocorm Like Bodies (PLB) induction from shoot tip of V. sumatrana, 2) knowing the PLB response of V. sumatrana to concentrations and soak period of colchicine and 3) find the effective concentrations and soak period of colchicine to induce tetraploid on PLB of V. sumatrana. Shoot tips from in-vitro cultured of V. sumatrana were subcultured on Murashinge and Skoog (MS) medium supplement with 3 mg/l BAP + 0,5 mg/l NAA, 3 mg/l BAP and 1,5 mg/l BAP. PLB of diploid V. sumatrana from the best treatment were soaked in 0.05% and 0.1% colchicine for 24 and 48 hours respectively in MS liquid medium, as control were set PLB without colchicine treatment. The results showed that MS medium supplemented with 1.5 mg/l BAP was the best formula to induce PLB. The highest percentage of survival rate of PLB and percentage of survived PLB regenerated shoot was obtained from 0.05% colchicine with 24 hours soak period treatment. The effective treatment to induce tetraploid on PLB of V. sumatrana Schltr. was obtained from 0.05% colchicine solution for 24 hours soak period.

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