dissertation

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EURASIAN NATIONAL UNIVERSITY named
after L.N.Gumilov
Effect of auxin, cytokinin and nitrogen on
anthocyanin
biosynthesis in callus cultures of redfleshed apple
Performed by: Ablayeva Akzharkyn
Astana 2017
Different from domesticated apples, red-fleshed apples possess a
dramatically high anthocyanin phenotype with highly pigmented
foliage, floral, and fruit tissues.They are very important genetic
resources to breed new cultivars characterized by red fruit flesh,
which are more attractive to the consumer and that promise a
health-promoting effect in human diet due to anthocyanins.
Callus culture is an effective and safe approach to produce
pigments, such as anthocyanins, on a large scale from plants to
meet the increased demand for natural food additives instead of
chemical synthesis. Anthocyanins also have antiinflammatory
activity, promote visual acuity, and hinder obesity and diabetes.
to investigate the effect of auxin alone and
auxin combined with cytokinin or nitrogen deficiency on
anthocyanin synthesis.
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1) to study the induction and acquisition of red and yellow calli;
2) to investigate the calli growth and dynamic levels of anthocyanins;
3) to compare transcript levels of anthocyanin pathway genes in red
and yellow calli;
4) to investigate the effect of auxins in suppression of anthocyanin synthesis;
5) to study the interaction of auxins with cytokinins in anthocyanin
biosynthesis;
6) to reverse the inhibitory effect of auxins on anthocyanin synthesis by
nitrogen deficiency
Methods of study
 cultura methods;
 determination of callus growth;
 extraction of anthocyanin and absorbance measurement;
 PCR amplification of the R6 region in the MYB10 promoter;
 RNA isolation, semi-quantitative and real-time RT-PCR;
 auxin treatments;
 cytokinin treatments;
 statistical analysis.
Red-fleshed apple individual which
was the hybrid offspring of Malus sieversii f.niedzwetzkyana
and ‘Fuji’.
Fig. 1 The foliage, flower and fruit of a R6/R6 homozygous individual redfleshed apple that was the hybrid offspring of M. sieversii f. niedzwetzkyana
and ‘Fuji’ used to induce calli and the PCR identification of its MYB10
promoter R6 segment (Lane 1), water as negative control (Lane 2). Calli
induced from leaves were red but turned yellow on part selective media.
Figure 2. a Red calli on optimal subculture medium supplemented
with 0.3 mg/L NAA and 1.0 mg/L BAP. b Yellow calli on optimal
subculture medium supplemented with 0.6 mg/L 2,4-D and 0.5
mg/L TDZ. c Yellow calli transferred back onto medium
supplemented with 0.3 mg/L NAA and 1.0 mg/L BAP. Green bar
indicates 0.5 cm.
Fig. 3 Red calli cultured 15 days on the medium supplemented with 0.6 mg/L NAA
and 0.5 mg/L BAP were transferred onto MS media respectively supplemented with
(1) 0.5 mg/L TDZ, (2) 0.6 mg/L 2,4-D, (3) 0.6 mg/L NAA combined with 0.5 mg/L
TDZ, and (4) 0.6 mg/L 2,4- D combined with 0.5 mg/L BAP. Red calli on the
medium supplemented with 0.6 mg/L NAA and 0.5 mg/L BAP as control (0). After 18
days, the anthocyanin accumulation was measured.
Fig. 4 A 21-day time course of calli growth (a) and anthocyanin content (b) of red and
yellow calli. The 15-day-old (after inoculation) red calli (0.3 g) and yellow calli (0.3
g) were cultured on their optimal subculture media and then harvested at days 0, 3, 6,
9, 12, 15, 18 and 21. The level of anthocyanins is represented by the absorbance
values measured at 530 nm divided by the sample’s fresh weight (Abs/g FW).
CONCLUSION
1. Auxin alone significantly inhibited anthocyanin
biosynthesis by suppressing the expression of
anthocyanin regulatory genes (MdMYB10 and
MdbHLH3) and structural genes.
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2. Co-treatment of auxin and cytokinin (BAP or
TDZ) significantly enhanced the cytokinin induced
increase in anthocyanin levels but auxin at high
concentrations strongly inhibited anthocyanin synthesis
even in the presence of cytokinin.
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3. Nitrogen deficiency could reverse the inhibition of
anthocyanin synthesis by auxins.
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The expert in the field of
biotechnology, selection, molecular
plant biology. The author more than
170 scientific publications, 10
innovative patents of the Republic of
Kazakhstan, the patent of the
Russian Federation, potatoes grade
"Astanalyk", sorts of wheat "Kazakhstan 20", and " Ak Orda".
The owner of the State scientific
scholarship for the scientists and
experts who have made an
outstanding
contribution
to
development
of
science
and
technology (2014).
Main publications
• Слугина М.А., Борис К.В., Какимжанова А.A., Кочиева Е.З.
• Внутривидовой полиморфизм генов сахарозосинтазы картофеля
сортов российской и казахстанской селекции Генетика. 2014. Том 50,
№ 6, - с. 677-682. Impact Factor 0,427.
• Какимжанова А.А., Каржауов М.Р., Каримова В.К., Раманкулов
Е.М.
• Генотипирование картофеля на основе RAPD-маркеров //
Биотехнология. Теория и практика. №1, 2011, - С. 32-38.
• Какимжанова А.А., Каримова В.К., Каржауов М.Р. и др.
• Использование молекулярно-генетических маркеров для идентификации
картофеля // Вестник, КазНУ, Серия биологическая, №2 (48) часть
2, 2011, - С. 237-239.
• Kakimzhanova A., Karimova V., Magzumova G. et al.
• Creating valuable forms and varieties of potatoes that are resistant to fungal
diseases // Current Opinion in Biotechnology // - 2013. - Vol. 24, Supplement
1, - P. 125. Impact Factor 7,860.
• Turganbayeva A., Hapilina O., Shek G. et al.
• Development important forms of soft spring wheat in North Kazakhstan //
Current Opinion in Biotechnology. 2013. Volume 24, Supplement 1, Pages
S.127. Impact Factor 7,860.
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Uvarova E.A., Belavin P.A., Kakimzhanova A.A. et al.
Oral Immunogenicity of Plant-Made Mycobacterium tuberculosis ESAT6 and CFP10 // BioMed
Research International. Volume 2013 (2013), Article ID 316304, 8 pages. Impact Factor 2,880.
Турганбаева А.К., Какимжанова А.А., Ергалиева А.Ж. и др.
Инновационный патент № 75334 от 07.11.2011 г. «Набор синтетических
олигонуклеотидов для выявления ДНК генномодифицированного рапса линии MS8, RF3
и RT73 методом полимеразной цепной реакции в режиме реального времени».
Белавин П.А., Какимжанова А.А., Раманкулов Е.М. и др.
Патент Российской Федерации на изобретение №2468082 от 27.11.2012.
«Рекомбинантная плазмида pBi121-ESAT6-GFP10-gIGN, кодирующая белки ESAT6 и
GFP10 Mycobacterium tuberculosis и φ-инерферон человека в трансгенных растениях»
Какимжанова А.А., Раманкулов Е.М., Каримова В.К. и др.
Положительное решение № 2011/095.4 от 29.11.2011 г. на новый сорт картофеля
Астаналық.
Шек Г.О., Хапилина О.Н., Турганбаева А.К. и др.
Положительное решение № 2011/096.4 от 29.11.2011 г. на новый сорт пшеницы «Ақ
Орда».
Шек Г.О., Турганбаева А.К., Какимжанова А.А. и др.
Положительное решение № 2011/097.4 от 29.11.2011 г. на новый сорт пшеницы
«Казахстан 20».
Али А.М., Какимжанова А.А., Каримова В.К. и др.
Инновационный патент РК №22559 от 26.04.2010 г. «Способ получения безвирусных
миниклубней картофеля на аэропонной установке»
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