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Development of tolerant/resistant cotton crop plants by using the applications of biotechnology against the adverse effects of climate


Description

Summary

Agricultural output is closely linked with the supply of irrigation water. Cotton being a cash crop contributes significantly in foreign exchange earnings. Production of cotton was less than the target mainly due to a number of factors including shortage of irrigation water. Decreasing ground water supplies and high energy cost affect production of irrigated cotton. In this regards we need such cotton varieties that tolerate the drought conditions. Some plants have the inherent ability to cope with environmental stresses through complex mechanisms which may induce similar cell signaling pathways (Kosmas et al 2006). Breeding for climatic stress tolerance through classical means has not been successful mainly due to the fact that this trait is multi-genic, requiring a coordinated action of several genes (Wang et al 2003). Attempts have been made to generate transgenic plants, harbouring one or two genes involved in the complex stress-countering pathways. Such plant transformations result in marginal stress tolerance because these manipulations affect only one stop in the complex cascade of multi-step pathways (Cherian et al 2006).  

Asiatic cotton Gossyium arboreum has built-in desirable genes for environmental stresses and resistance to insect pests.  It is important in genomic studies by sharing it’s A genome to modern upland cotton, G. hirsutum. Its diploid nature of gnome made it right choice for identification of novel genes in genus Gossypium.

Above in view, it is proposed to isolate genes from local cotton varities of Gossypium arboreum which is highly stress tolerant and to transform these genes to sister cotton plants Gossypium hirsutum which are high yielding with better fibre quality but susceptible to abiotic stresses. It is anticipated that it will yield better quality and stress tolerant cotton plants. This will further explore for other agriculturally important crop plants to make them compatible with climate change threats.


Is this proposal for a practice or a project?

Project


What actions do you propose?

Main objective

This project shall focus on the identification of drought related genes in Gossypium arboreum specie of cotton and those identified genes in future can be used to engineer the drought tolerance in American cotton and other agriculturally important transgenic crops with improved character of drought tolerance. We are already working with the drought related studies in CEMB and different local desi cotton varieties are under investigation. Presently in Pakistan, there are 52 approved varieties of upland cotton, 6 Desi, 8 Bt and one Bt hybrid but none of the cotton variety is approved for drought tolerance (Central Cotton Research Institute, Multan, Pakistan, personal communication. Therefore for control studies there is none of the known cotton variety for drought tolerance, so for control studies purpose, the same variety in well watered condition will be used as the control.

Specific Tasks

-Plantation of local Gossypium arboreum cotton plants in the field

  • Soil/land will be prepared according to the recommended agricultural practices such as fertilizer, irrigation etc 
  • Local varieties of cotton (Gossypium arboreum) will be grown in the CEMB green house 

 

-Identification & isolation of drought tolerant genes from local  cotton plants Gossypium arboreum

  • Drought application will be given to the plants by with- holding the irrigation.
  • Leaf/root samples will be taken from the water stressed plants and stored in the liquid nitrogen
  • Total RNA will be extracted from the samples by the standard protocols and reverse transcribed to maake cDNA.
  • Oligonucleotide Primers will be designed  
  • Full length genes showing homology with drought tolerant transcripts will be isolated and characterized using RACE and genomic DNA PCR.
  • The gene expression will be verified for confirmation of the transcript.
  • Cloning of isolated genes into bacterial expression vectors and marker/reporter gene will be attached for transient expression studies of the transgene into the transgenic crop plants 

 

-Transformation of these genes into Gossypium hirsutum

  • Construct of  drought tolerant genes will be made into plant expression vector pCAMBIA 1301 and transformed into local variety of G. hirsutum through Agrobacterium mediated transformation and drought tolerant plants will be developed.
  • Selection of transformed plants on the antibiotic selection medium
  • Development of roots of the transformed plants
  • Hardening off transgenic plants in the soil  

 

-Molecular analysis of transgenic plants and screening of transgenic plants on the basis of Bioassay

  • Positive plants are confirmed through PCR and Gus assay, real time PCR, bioassays Southern blot, Western Blot etc

-Field studies of transgenic plants

  • Seeds of selected transgenic plants will be germinated under controlled conditions and will be subjected to field conditions in the CEMB fields. Plant’s conditions will be subjected to drought stress by controlled irrigation at vegetative, flowering and boll formation stages. Control or well watered non transgenic plants will also be planted for comparisons. Biosafety guidelines will be followed to the field trial.
  • Morphological, biochemical and physiological analyses of transgenic plants will be done to asses the behavior of transgenic plants under the environmental challenges 

 

-Development of genetically modified plant variety

  • Breeding technology will be used for the development of approval of the local genetically modified cotton variety
  • Different local seed companies will participate in the project to develop the transgenic cotton crop as genetically modified variety approved from national seed council and environmental protection agency.
  • Plant will be tested according to the biosafety guidelines
  • Biosafety commission will be contacted for approval of the biosafety guidelines. Seed production and multiplication will be done at different localities of cotton growing areas.
  • Crop testing will be done at different local cotton growing areas specially the barren lands to see the effects of water shortage on the crop
  • Dissemination of knowledge and information at the national and international level for adoption of technology by other stake holders.
  • Farmers form the local cotton growing areas will be invited to show the technology results and to convince them to adapt the new eras to overcome the losses to crops by environmental threats. 

 


Who will take these actions?

Centre of Excellence in Molecular Biology, University of The Punjab Lahore, 87 West Canal Bank Road Thokar Noaz Baig Lahore, Pakistan is using the similar technology.

We have isolated two stress proteins from Gossypium arboreum designated as GUSP1 and GHSP26. We have also characterized two transcriptional factors of these drought tolerant genes and identified them as  (1) Zinc finger protein gene transcription factor that binds to   stress responsive (MYB binding sequence) cis-acting element of GUSP promoter and (2) Homeodomain of leucin zipper transcriptional factor that bind to abscisic acid responsive element (ABRE) of the GHSP26 promoter. We hypothesize that, when transformed these genes into different plant species will yield drought tolerant plants.  We have transformed these genes into local crop varieties and generated transgenic plants. Our initial studies are very encouraging. Preliminary data show that transgenic plants have acquired inherent abilities to cope with environmental stresses as compared with parent cotton plants.

The owner of the project Dr Bushra Rashid has enough experience in plant biotechnology and plant genomics. She has a group of scientists including faculty members and PhD research scholars having expertise in the relevant field.  This research work is proposed to be carried out under her supervision in plant  genomics lab at above mentioned institution. The dedicated team members will actively participate in the development of materials proposed in this project 

Private sector seed companies will be involved for field trials of locally developed transgenic plants and multiplication of transgenic seeds to be supplied to the stake holders.

Local farmers will also be invited to take part in the growing of transgenic crop at their local farms to see the benefits of the technology.

Government organizations such as national biosafety commission, national seed council and ministry of environment  and other relevant local government officials may also be invited to have the relevant certification for the approval of technology.

International organizations such as Monsanto, Bayer, DuPont and other agriculture based industries may also be collaborated to compare the technology results and adoption of our locally developed materials at international level.

Information related to the technology will be disseminated through organizing the national and international  conference, symposia and training workshops specially to the participant from under developed countries.  Local media will be invited to disseminate the achievements' information at the national level. International collaborators may also be invited to disseminate the technology specially to the low income and technologically far behind countries.       


Where will these actions be taken?

These actions will be taken at Centre of Excellence in Molecular Biology, University of The Punjab Lahore, 87 West Canal Bank Road Thokar Noaz Baig Lahore, Pakistan. Moreover local cotton growing areas will be used to grow the locally developed cotton crop which is proposed to adapted for climatic change.   


In addition, specify the country or countries where these actions will be taken.

Pakistan


Country 2

No country selected


Country 3

No country selected


Country 4

No country selected


Country 5

No country selected


Impact/Benefits


What impact will these actions have on greenhouse gas emissions and/or adapting to climate change?

The project outcomes will produce the drought tolerant /resistant  cotton crop. This will be helpful to cultivate the crop on rainfed areas and where the irrigation water is less.

Social benefits of project outcomes (with special reference to increase in export, decrease in import, employment generation, environmental protection, health improvement, improvement in food quality

There may be increase in employment generation as the yield is expected to increase 8% as well as increase in oil production so to manage this increase employment generation is expected in the fields, oil factories and ginning/textile mills etc. It is expected that due to Bt cotton varieties there will be less pesticides use and it is environmentally safe technology. Moreover there will be saving of irrigation water so the project is expected to save water for other irrigation purposes and even technology could be more benefited for use in barani or less irrigation areas.

Benefit to the poor or small farmers/stakeholders

The limiting factor for small/poor farmers is that he has limited resources and he may not have too many choices. So by spending just Rs.600/kg extra on cost of seed it is expected to save Rs 10000/acre for the cost irrigation water and more than half/acre of the pesticide use and the farmer can get the benefit of 8% increase of the yield. After the approval of variety from the commission it will take maximum three to five years that the 30% end user will adopt the technology

Requirements of project technologies for adoption

The ultimate target for marketing of crop biotechnological products are the farmers, so development of cotton crop with enhanced drought tolerance will help the community to get improved yield especially in barani or water shortage areas. The project technologies for adoption are not difficult and normal procedures for commercialization or publicity will be used for adoption of this technology.

EXPECTED RESULTS

  • Drought tolerance from Gossypium arboreum will be isolated.
  • These genes will be transformed into choice host plant.
  • Drought tolerant plants will be developed.
  • Research publications will be submitted in peer review Journal.
  • Manpower will be trained in latest techniques of molecular biology which can prove as asset to work in other projects for participating laboratories.
  • Drought tolerant/resistant cotton seeds will be available to the farmers which will save input use and earn the extra benefit

 


What are other key benefits?

A- Economic benefits of the project through:

a- Yield improvement

1.Current yield of cotton in Pakistan = 1711 kg/ acre

(Reference Economic Survey of Pakistan)

2. Total Current Area of Cotton in Pakistan= 6993600 acres

3. Price per kg=Rs 57/kg

4. Increased promised yield=23kg/acre (during drought)

i. Value= Promised yield improvement (kg/acre)/2 xTotal current area in acres/3 xCurrent Price (Rs/kg)/1000000

23/2 x 6993600/3 x 57/1000000 =1528.10 (Million Rs)

ii. Additional cost for increased yield:

For Bt/drought tolerant cotton seeds we expect an increase in the cost of seeds @ the cost of Rs 600/Kg, so

Expected increase in cost (Rs/acre)/1000000 x Current area in acres/3

600/1000000 x  6993600/3 = 4196 Rs

iii. Net Value (Rs. M)                     (i) – (ii)

          1528.10 – 0.0041 = 1528.09 M Rs

B. Decrease in input use

1- For Drought tolerant technology

  1. Normally cotton requires irrigations in total =15 (1 irrigation costs Rs 2000/ acre)
  2. Through drought tolerant technology saving of 05 irrigations= Rs 10000/acre will be saved, Therefore; 

Value (Rs M) = Promised decrease in input use (Rs/acre) x Current area (acres) /2x1000000 x3

10000 X 6993600/ 2x 1000000x 3 = 2331.0 Rs M

Reference for these calculations

  • Denise McWilliams 2003
  • Ihsan Ullah, Mehboob-ur-Rahman, Muhammad Ashraf, Yusuf Zafar 2008.
  • Sulian Lv, Aifang Yang, Kewei Zhang, Lei Wang, Juren Zhang. 2007 I
  • Monsanto ~ R&D pipeline

 

2. Benefit For using Bt Transgenic seedS

1.Current Number of insecticide sprays required for cotton crop = 8- 9 sprays @ of Rs.500/spray

2.Total cost of spraying  = 500 & 9   = Rs 4500

3.Potential saving of sprays =2.73 times

Value (Rs. M) = 1369 (Rs. Per acre) X 6993600 acres/2x1000000 x 3 = 1595 M Rs

Reference for calculation: (Prospects for Bt Cotton Technology in India, R. B Barwala, V. R Gadwai. Usha Zafar, Brebt Zehr, AgBIoforum Volume 7) 

Net value =  2331+1595 = 3926 M Rs / annum

Other benefits (Rs M)), basis and formula of calculation

  1. Total cotton seed oil produced presently in Tonnes= 371,000 (Reference Economic Survey of Pakistan 2008-09, Chapter#2 Agriculture Table 2.9)
  2. Expected increase in cotton seed oil due to 8%  increased yield in Tonnes= 29680
  3. Cotton seed oil price/kg Rs =100/kg
  4. Total value increase Rs (M) =2.968

 

This is a two way project i.e. combination of drought tolerance and Bt technology we will have two benefits. We are expecting increase in yield of cotton because of drought tolerance and control of pests and  we will be saving use of irrigation water and pesticides. So this  technology will  be

  1. Increasing farm income
  2. Reducing input use
  3. Saving foreign exchange , Currently we are spending roughly 220M$US annually on import of pesticides and out OF this, around 72% (158 M US$)  is used on cotton, so a potential annual saving of 47 M US$ in terms of foreign exchange spending. 


Costs/Challenges


What are the proposal’s projected costs?

The project cost is the estimated one at this time. The research work taken will have the estimated cost. Therefore, the estimated cost of the actions taken is proposed to be US$ 110000 which includes the inputs, consumables, equipments, salaries and infrastructures.

The challenges for the implementation of this project is the academia-industry linkage. It is challenging to convince the seed companies to participate or collaborate for the adoption of new transgenic/ genetically modified crop technology in comparison of the old classical breeding methods.

It is also challenging to invite the farmers especially the small land farmers to adapt the new technology as they are strict to the old methods of farming. But the field trials at different locations nearby their vicinity will help them out to attract them to see the performance of the crop developed by the advanced applications of biotechnology       


Timeline

This proposal will have the impact in the short term 1-15 years. The proposal is expected to start by early 2018 and the first phase of basic research will be completed after 5 years. The second phase of applied research will be done/complete after 10 years of the start date and the commercial application will be implemented after completion of 15 years and then this will be a routine practice that the outcomes of the project will be available to the nation.

1st phase of the project: The basic research (0-5 years)

Development of transgenic crop technology at the proposed institution and testing of crop at the laboratory level  

2nd phase of the project: The applied part of the project (5-10 years)

The transgenic crops' seeds will be given to the collaborating seed companies for field trials at different locations of cotton growing areas. The low irrigation areas and the rainfed areas will also be chosen to find the crop testing for growing in water shortage areas. Data will be collected and analyzed  

3rd phase of the project: The commercial application of the research (10-15 years)

The growing of transgenic cotton crop throughout the country at different localities at commercial level and the data for the expected outcome will be analyzed. Approval of the crop as variety will be sought from the relevant bodies/organizations for certification and approval. Technology will be disseminated through the media and the the collaborating companies will be invited for conferences, symposia and the technology transfer agreements.  


About the author(s)

The owner of the project Dr Bushra Rashid has enough experience in plant biotechnology and plant genomics. She has a group of scientists including faculty members and PhD research scholars having expertise in the relevant field.  This research work is proposed to be carried out under her supervision in plant  genomics lab at above mentioned institution. The dedicated team members will actively participate in the development of materials proposed in this project. Dr Bushra Rashid has completed other research projects granted by Higher Education Commission Pakistan. The prof of her achievements has been documented as research publications, book chapters, review articles, full length gene sequences submitted at PUBMED etc. She has quite good experience to work in the international labs such as University of Edinburgh, Scotland, UK, Commonwelath Scientific and Industrial Research Organization, Canberra, Australia, University of Illinois, Urbana-Champaign, USA and Harran University, Sanliurfa, Turkey  


Related Proposals

No other proposal of this sort


References

  • Rivero RM, Kojima M, Gepstein A, Sakakibara H, Mittler R, Gepstein S, Blumwald S. 2007. Delayed leaf senescence induces extreme drought tolerance in a flowering plant. Proc Natl Acad Sci USA 104(49): 19631-6.
  • Sawahel WA. 2001. Stable Genetic Transformation of Cotton Plants Using Polybrene-Spermidine Treatment.  Plant Mol Biol Rep19: 377a–377f.
  • Somers DA, Makarevitch I. 2004. Transgene integration in plants: poking or patching holes in promiscuous genomes? Curr Opin Biotechnol 15(2):126–131.
  • Latham JR, Wilson AK, Steinbrecher RA 2006. The Mutational Consequences of Plant Transformation. J Biomed Biotechnol 1–7.
  • Shinozaki K, Yamaguchi-Shinozaki K, Seki M. 2003. Regulatory network of gene expression in the drought and cold stress responses. Curr Opin Plant Biol 6 410–417
  • Kosmas SA, Argyrokastritis A, Loukas MG, Eliopoulos E, Tsakas S, Kaltsikes PJ 2006. Isolation and characterization of drought-related trehalose6 phosphate synthase gene from cotton (Gossypium hirsutum L). Planta 223:329-339
  • Wang W, Vinocur B, Altman A 2003. Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta  218:1-14
  • Cherian S, Reddy MP, Ferreira RB 2006.Transgenic plants with improved dehydration-stress tolerance: progress and future prospects.  BIOLOG. PLANTA.50 (4): 481-495.
  • Denise McWilliams 2003. Drought strategies for cotton. Cooperative Extension Service Circular 582, College of Agriculture and Home Economics, New Mexico State University NMSU and the U.S. Department of Agriculture cooperating. nmsu.edu/pubs/_circulars/CR582.pdf
  • Ihsan Ullah, Mehboob-ur-Rahman, Muhammad Ashraf, Yusuf Zafar 2008.  Genotypic variation for drought tolerance in cotton (Gossypium hirsutum L.): Leaf gas exchange and productivity Flora 203:105–115
  • Sulian Lv, Aifang Yang, Kewei Zhang, Lei Wang, Juren Zhang. 2007 Increase of Glycinebetaine synthesis improves drought tolerance in cotton Mol Breeding 20:233–248
  • Monsanto ~ R&D pipeline ~ cotton pipeline products (http://www.monsanto.com/products/pipeline/cotton/products.asp)