Search or add a thesis

Advanced Search (Beta)
Home > Genetic Transformation and Expression Analysis of Cold Tolerant Gene in Tomato Solanum Lycopersicum Mill.

Genetic Transformation and Expression Analysis of Cold Tolerant Gene in Tomato Solanum Lycopersicum Mill.

Thesis Info

Access Option

External Link

Author

Shah, Sabir Hussain

Program

PhD

Institute

The University of Agriculture

City

Peshawar

Province

KPK

Country

Pakistan

Thesis Completing Year

2015

Thesis Completion Status

Completed

Subject

Botany

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/6905/1/Sabir_Hussain_Shah_2015_Plant_Biotechnology_IndegenScholar_UA_Peshawar.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726314313

Asian Research Index Whatsapp Chanel
Asian Research Index Whatsapp Chanel

Join our Whatsapp Channel to get regular updates.

Similar


Chilling stress severely reduces the productivity of tomato as it is a cold sensitive plant. CBF3/DREB1A plays a key role in generating cold tolerance in tomato by regulating the response of multiple genes under chilling stress. In this study, cold tolerant gene (DREB1A) driven by Lip9 promoter, was transformed in three tomato genotypes (Rio Grande, Moneymaker and Roma) through Agrobacterium tumefaciens, employing tissue culture dependent and tissue culture independent transformation strategies. For tissue culture dependent transformation strategy, the effects of various PGRs (IAA, NAA, ZEA, Kin, BAP and GA3) and two ethylene inhibitors (AgNO3 and CoCl2) were investigated on callus induction and in vitro shoot regeneration. The maximum callus induction frequency (67.48%) was recorded on MS basal media enriched with 2.0 mg/l IAA, 2.5 mg/l BAP in cv. Rio Grande followed by Roma (62%) and Moneymaker (58.23%). Supplementation of AgNO3 (10-15 mg/l) in MS basal media along with PGRs (2.0 mg/l IAA, 2.5 mg/l BAP) significantly yielded the highest callus induction frequency (91.83%) in cv. Rio Grande, followed by Moneymaker (82.66%) and Roma (88.33%). Similarly, in vitro shoot regeneration frequency on MS media fortified with 0.1 mg/l IAA, 1.0 mg/l ZEA and 2.0 mg/l BAP significantly enhanced with the addition of 8-10 mg/l AgNO3 in all the cultivars i.e. in cv. Rio Grande (96.65%) followed by Roma (92.66%) and Moneymaker (90%). Likewise, the highest callogenesis (75.65%) was recorded in cv. Rio Grande on callus induction medium (CIM) supplemented with CoCl2 (5.5 mg/l), IAA (2.0 mg/l) and BAP (2.5 mg/l) followed by cv. Roma whose best callus induction (73.66%) was obtained on CIM supplemented with CoCl2 (4.5 mg/l), IAA (1.0 mg/l) and BAP (2.5 mg/l). In case of cv. Moneymaker the best callogenesis (68%) was secured on CIM having CoCl2 (3.5 mg/l), NAA (2.0 mg/l) and BAP (2.0 mg/l). The xiv highest in vitro shoot regeneration (85%, 81% and 78%) was recorded in Rio Grande, Moneymaker and Roma, respectively on shoot induction media supplemented with CoCl2 (4.25 – 5.0 mg/l). During this study, various concentrations of sucrose and sorbitol were scrutinized on in vitro shoot culture. The highest in vitro shoot regeneration frequency (100, 97.69 and 99%) was recorded in Rio Grande, Moneymaker and Roma with accumulative effect of sucrose and sorbitol (30: 30 g/l). Subsequently, transformation experiments were conducted by optimizing various factors both for tissue culture based and in planta techniques. For tissue culture based method of transformation; fifteen days old in vitro seedlings, forty-eight hours pre-culture period, bacterial density (OD600 nm = 0.2), three minutes infection period, 60 μM acetosyringone, forty-eight hours co-cultivation period, pH 5.6 of co-cultivation media, six days pre-selection duration, cefotaxime (500 mg/l) and hygromycin (35 mg/l) as lethal dose were found optimum. For in planta technique of transformation, various factors such as growing medium; soil: vermiculite (1: 1), optical density (OD600 nm = 1.0) and incubation period (20 min) were found optimum for efficient transformation efficiency. Polymerase chain reaction, Multiplex polymerase chain reaction, Southern blotting and Reverse transcriptase PCR confirmed the presence, integration and expression of DREB1A in T0 - T2 transgenic lines. Physiological and biochemical analyses of T2 transgenic plants depicted that after various chilling stresses; stomatal conductance, transpiration rate, CO2 concentration rate, photosynthetic rate, relative water contents, proline contents, total soluble sugar contents, chlorophyll contents, carotenoid contents and ascorbic acid contents of transgenic lines were significantly higher than those of NT plants. These findings clearly indicate that transgenic tomato plants over-expressing Arabidopsis CBF3 gene enhanced protection and provided cold tolerance under controlled conditions in transgenic containment.
Loading...
Loading...

Similar Books

Loading...

Similar Chapters

Loading...

Similar News

Loading...

Similar Articles

Loading...

Similar Article Headings

Loading...

ٹریفک کے قوانین

ٹریفک قوانین
نحمدہ ونصلی علی رسولہ الکریم امّا بعد فاعوذ بااللہ من الشیطن الرجیم
بسم اللہ الرحمن الرحیم
معززصدرو میرے ہم مکتب سا تھیو!
آج مجھے جس موضوع پرلب کشائی کرنی ہے وہ ہے:’’ٹریفک قوانین‘‘
صدرِذی وقار!
قانون، اصول، ضابطہ جیسے الفاظ کا تصور جب دماغ کے در پچوں کو دستک دیتا ہے تو تہذیب وتمدن ، اورمنظم قوم کی ایک تصویر بھی دماغ کے خانوں میں ابھرتی ہوئی محسوس ہوتی ہے، اور اُجلا پن جگہ جگہ دکھائی دینا شروع ہو جا تا ہے۔ کائنات رنگ و بو میں ہر شے کا اپنا اپناضابطہ ہے۔ نظام ِشمسی ہو، نظامِ فلکی ہو ،نظامِ ارضی و سماوی ہو، جملہ نظام ہائے حیات قوانین کے دائرے میں متحرک نظر آتے ہیں۔ کچھ قوانین ایسے ہوتے ہیں جن کو انسان اپنی بقاء کے لیے بناتے ہیں، انہی قوانین میں ٹریفک کے قوانین بھی ہیں۔
صد رِمحترم!
زمین پر حشرات الارض کو دیکھیں تو ان کی اجتماعی حرکت ایک قطار میں نظر آئے گی۔ آسمان کی بلندیوں پرمحوپرواز طائران خوش الحان کی زندگی کا مشاہدہ کریں تو ان کی پرواز بھی کسی قانون اور ضابطے کے تحت ہوگی۔ حدی خواں کے اونٹوں کی قطاریں، بلبل کی چہک، پھول کی مہک ، جگنو کی چمک، ستاروں کی دمک ، سورج کی روشنی، چاند کی چاندنی ، فضاؤں کی سرسراہٹ ، آبشاروں کی گڑگڑاہٹ ، سمندر کا سکوت، دریا کا شور، صبح سہانی ، ندیوں کی روانی یہ جملہ مظاہرِ فطرت کسی نہ کسی ضابطے کے تحت سرگرم عمل ہیں۔
معززصدر!
قوانین انسان کی فلاح کے لیے بنائے جاتے ہیں، انسان کی ترقی مقصود ہوتی ہے، انسان کی زندگی میں حسن پیدا کرنا ہوتا ہے، انسان کونشست و برخاست کا ڈھنگ سکھانا ہوتا ہے، انسان کی گفتار میں شائستگی پیدا کرنا ہوتی ہے۔ انسان کی رفتار میں اعتدال پیدا...

سکھ دھرم پر اسلامی تعلیمات کے اثرات کا علمی جائزہ

Sikhism is one of the Non Semitic religions founded by Guru Nanak, belonged to a Hindu family and was born in 1469 A.D. This religion is popular in India and Pakistan. Some inhuman customs in Hinduism like caste system, the custom of Satty (burning out of wife with the dead body of husband), monopoly of Brāchman etc. Compelled him to introduce a new religion based on equality and justice. As identified from the life style of the founder of Sikhism and his followers, he is deeply impressed with Islamic teachings. Their habits and customs reflect an Islamic picture. Guru Nanak was a monotheist and was against the worship of idols. He believed in equality and acknowledged the prophet Muhammad (S.A.W), as a role model for human beings. This article is aiming to explain the teachings of Sikhism derived from Islam.

Evaluation of Different Stress Indicators Associated With Repproductive Cyclicity of Crossbred Dairy Cows

Eminent environmental temperature has adverse affect on dairy cattle production and reproduction. The periparturient period is also very critical for reproductive functions of dairy cows. A variety of methods are used to improve fertility in these stress conditions. The present study was conducted under a series of three experiments to find out stress-reproduction relationship and effect of vitamin E on this interaction. Study-I, was conducted to assess physiological and reproductive changes, in relation to heat stress in different dairy cattle breeds in various seasons. A total of thirty six lactating dairy cows, nine from each breed of local (Sahiwal and Achai), Cross-bred (Sahiwal x HF) and Holstein Friesian were selected. Sampling was conducted when the cows were in the di-estrus phase of the estrus cycle in February (thermoneutral), April (thermotransitional) and June (thermal stress) at an average environmental temperature of 18°C, 32°C and 42°C respectively. A significant decrease in Daily milk yield (DMY) was observed in all breeds with increase in environmental temperature (P < 0.01). DMY also varied significantly (P < 0.001) among breeds. The HF dairy cows showed the highest mean DMY followed by cross-bred, Sahiwal and Achai at all the three seasons. BCS was not affected by environmental temperature; however, BCS varied among breeds significantly (P < 0.01). A significant increase in all the physiological parameters (rectal temperature, respiratory rate and pulse rate) was observed in all dairy cows with increase in environmental temperature during thermo transitional and summer seasons in April and June respectively. Thermal stress in June increased concentrations of glucose (P= 0.014), cortisol (P < 0.001), heat shock protein-70 (HSP-70) (P < 0.001) and follicular numbers (P < 0.01), while decreased progesterone concentrations (P < 0.01) in all four breeds as compared to February. DMY and progesterone level are negatively correlated with environmental temperature. Glucose and cortisol levels were positively correlated with each other and negative with Progesterone. Positive correlation was found between glucose, cortisol and HSP-70 with rise in environmental temperature. However progesterone was negatively correlated with these parameters at high environmental temperature. It is concluded that the intensity of changes was higher in HF and cross-bred dairy cows than local Achai and Sahiwal, suggesting that extra managemental and therapeutical support is needed to improve fertility of these breeds in tropical regions. Study-II, was designed to detect the occurrence of oxidative stress in dairy cows by determining the plasma malondialdehyde (MDA) concentration during the periparturient period and its possible relationship with dairy cows fertility. Thirty-six periparturient lactating dairy cows, nine from each breed of Sahiwal, Achai, Cross-bred (Sahiwal x HF) and Holstein Frisian were selected. Sampling was conducted on post partum day (P.P.Day) 60, 82 and 105 of the lactation stage during the diestrus period of the estrus cycle. DMY varied significantly with P.P Days and among breed (P < 0.001). Highest DMY was shown by HF followed by cross-bred, Sahiwal and lowest by Achai. The interaction (PP.Days x Breed) effect was also highly significant (P < 0.01) for DMY. Serum glucose concentration significantly increased with increase in PP.Days (P < 0.01). Breed also affected glucose concentration (P < 0.001). A decrease in MDA concentrations were observed with increase in PP.Days (P < 0.001) from P.P day-60 to 82 and 105. Breed also affected the MDA concentrations significantly (P < 0.001), more prominent in HF and cross-bred. Both P.P. Days and breed significantly affected the serum cortisol concentration (P < 0.001). The cortisol concentration significantly decreased from P.P day-60 to 82 and 105. Progesterone level increased in all breeds (P < 0.05) with increase in P.P. Days (P < 0.001) from day-60 to 82, continued up to day-105. BCS was negatively correlated with serum cortisol, glucose and progesterone. Cortisol, DMY and MDA were positively correlated with each other and negative with progesterone and P.P. Days. The study revealed that HF and cross-bred dairy cows were more prone to oxidative stress than local breeds. Correlations analysis indicates that high milk production delayed ovarian activity effecting the reproduction of periparturient cows as assessed by progesterone concentrations. Therefore, it is suggested that higher genetic merit milk breeds should give extra managemental support for their better productivity and fertility. Study-III, was designed to find out the response of different breeds to vitamin E supplementation in combating stress and improving reproductive cyclicity. The experiment was conducted on thirty six lactating dairy cows, nine from each breed of Sahiwal, Achai, Cross-bred (Sahiwal x HF) and Holstein Frisian. Vitamin E was supplemented to all dairy cows in feed at a dose rate of 1000 IU/ cow/ day. Sampling was conducted on day-zero (control) before Vitamin E supplementation, day-20 and day-40 of Vitamin E supplementation during the diestrus phase of the estrus cycle. Daily milk yield and BCS varied significantly among breeds (P < 0.001) with no significant effect of vitamin E. Serum glucose increased significantly (P < 0.05) with vitamin E supplementation, but did not varied among breeds significantly. There was significant increase in SOD and GPx with vitamin E supplementation (P < 0.001). Breeds also varied in super oxide dismutase (SOD) and glutathione per oxidase (GPx) activities (P < 0.05, P < 0.001 respectively) with more increase in GPx in local dairy breeds. Stress markers, plasma MDA and HSP-70 and serum cortisol concentrations decreased significantly (P < 0.001) with vitamin E supplementation with significant variations among breeds (P < 0.001). Progesterone concentration increased significantly (P < 0.001) in all breeds with vitamin E. Breed effect on progesterone concentrations was also found significant (P < 0.05). Local breeds showed higher progesterone concentration while overall increase was found higher in Cross-bred dairy cows (39.95%). Number of dominant follicles decreased significantly with vitamin E supplementation (P < 0.01) in all the dairy breeds, highest in cross-bred dairy breeds. Breed also showed significant effect on number of follicles (P < 0.05). DMY showed positive correlation with MDA and negative with SOD, GPx and progesterone. The stress markers (cortisol, HSP-70 and MDA) were positively correlated with each other and glucose concentration while negative with antioxidant markers. Antioxidant markers were positively correlated with progesterone. It can be concluded that improved milk breeds showed more prominent response to vitamin E supplementation than local breeds suggesting regular feeding of antioxidant to these breeds for better performance.