Utilization of natural tools and biotechnology for clcv resistance in cotton

1. Utilization of Natural Resources
and Biotechnological Tools for
developing CLCuV Resistance in
Cotton
By Umair Rasool Azmi
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2. Outline
 Introduction and importance of cotton
 History and current status of CLCuV in Pakistan
 Mode of transmission and symptoms
 Role of weather in spreading of CLCuV
 Available sources and their utilization
 Biotechnological Tools for improvement
 References
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3. Introduction and Importance of Cotton
 Cotton is a fiber crop of global significance cultivated in more than 70
countries.
 Belongs to genus Gossypium which comprises about 50 species.
 4 are cultivated including 2 tetraploid (G. hirsutum & G. barbadense) and 2
diploid (G. arboreum & G. herbaceum).
 Statistics showed by FAO during the year 2017
 11.1% increase in cotton production across world.
 World total cotton production was around 25.6Mt (million tons).
(FAOSTAT, 2017)
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4. Cont…
 Status of cotton in Pakistan 2018-19
 0.8% share in GDP and 4.5% in agriculture value addition.
 Production of 9.86 million bales.
 Planted on an area of 2.37 million hectares.
(Pakistan Economic Survey 2018-19)
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5. History and Current Status of CLCuV in Pakistan
► History:
 1st time appeared in 1967.
 1st epidemic in 1993-94 caused 8.04 million bales reduction in yield.
 2nd epidemic in 2001.
(Mahmood et al., 2003)
► Current status:
 CLCuV causes about 30% yield losses annually.
(Hassan et al., 2016)
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6. Mode of Transmission
 The virus is obligately transmitted by silver leaf biotype of whitefly (Bemisia
tabaci).
 This vector allows the rapid and efficient replication of virus because it is an
indiscriminate feeder.
 Whitefly is responsible for 50% yield loss by reducing boll formation.
(Khan et al., 2015)
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7. Symptoms
Based on Severity Grade
Grade 0: No Symptoms
Grade 1. Blocking of Primary Veins Grade 2. Blocking of Secondary veins
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8. Cont…
Grade 3. Severe leaf curling and leaf
enations
Grade 4. Grade 3 symptoms + Stunted
growth
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9. Role of weather in Spreading CLCuV
 Several climatic conditions like rainfall, wind, humidity have a great
influence in the spread of the disease.
 Max. Temperature: 33-35.5°C
 Min. Temperature: 26-26.5°C
 Relative Humidity: 64-66%
 Rainfall: 1-2mm
 Windspeed: 5.5-5.7 Kmh-1
(Saeed et al., 2018)
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10. Available Sources and Their Utilization
► Germplasm
 G. arboreum (A2 Genome)
 G. anomalum (B1 Genome)
 G. thurberii (D1 Genome)
 G. armourianum (D2-1 Genome)
 G. tomentosum (AD3 Genome)
 Traits of Interest
 Presence of cuticular waxes.
 Presence of trichomes.
 Presence of oxalates and inorganic salts
(Azhar et al., 2010)
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11. Biotechnological Tools for Improvement
 Genetic Transformation
 Pre and Post Transcriptional changes
 TALEN
 RNAi
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12. Genetic Transformation
► Many transformation methods are available but in cotton two transformation
methods are mostly used
► Agro bacterium mediated
 A. tumefaciens
 A. rhizogenes
► Biolistics
 Gene gun
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13. Pre and Post Transcriptional Changes
 TALEN
 Modified Restriction Enzymes to cut specific Sequence(s).
 Discovered in Xanthomonas.
 Bind to promoter sequences and facilitate bacterial infections
 Success against CLCuV
 Transcriptional control of replication associated protein (Rep), encoded by AC1 of
geminiviruses at NIBGE, Faisalabad.
 Transformed Plants Showed high resistance level up to 81% against CLCuV.
(Hashmi et al., 2011)
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14. RNAi (Gene Silencing)
► Introduction of a double-stranded RNA (dsRNA)into an organism to induce
sequence-specific RNA interference (RNAi) of a target transcript.
► Success against CLCuV
 At National Agri-Food Biotechnology Institute, India VIGS-mediated RNAi was utilized to
affect the DNA of CLCV of Multan (CLCuMV).
 Such VIGS system showed efficient silencing of the target genes in
tobacco, Arabidopsis and Upland cotton plants.
(Kumar et al., 2014)
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15. References
 Azhar, M.T., M.U. Rehman, S. Aftab, Y. Zafar and S. Mansoor. 2010. Utilization of natural and
genetically engineered sources in Gossypium hirsutum for the development of tolerance against
cotton leaf curl disease and fiber characteristics. Int. J. Agric. Biol., 12, 744-748.
 Economic Survey of Pakistan, 2018-19
 Feehan, H.A.Q., W. Malik, A. Maqbool, M. Hassan, M.A. Rehman, M. Shoaib, M. Shehzad, S. Ahmad,
L. Ahmad and M. Arshad. 2016. Cotton Leaf Curl Virus (CLCuV) Disease in Pakistan: A Critical Review.
Appl. Sci. Bus. Econ. 3(1):8-14.
 Hashmi, J.A., Y. Zafar, M. Arshad, S. Mansoor and S. Asad. 2011. Engineering cotton (Gossypium
hirsutum L.) for resistance to cotton leaf curl disease using viral truncated AC1 DNA sequences. Virus
Genes. 42(2):286-96.
 Khan, M.A.U., A.A. Shahid, A.Q. Rao, N. Shahid, A. Latif, S. Din and T. Husnain. 2015. Defense
strategies of cotton against whitefly transmitted CLCuV and Begomoviruses. Adv. Life Sci. 2: 58-66.
 Kumar, J., S. Gunapati, J. Kumar, A. Kumari, A. Kumar and R. TulI. 2014. Virus-induced gene
silencing using a modified betasatellite: a potential candidate for functional genomics of
crops. Arch. Virol. 159, 2109–2113.
 Saeed, H., M. Ehetisham-ul-Haq, M. Atiq, M. Kamran, M. Idrees, S. Ali, M. Burhan, M. Mohsan, M.
Iqbal, S. Nazir and S. Il Yasin. 2018. Prediction of cotton leaf curl virus disease and its management
through resistant germplasm and bio-products. Arch. Phytopathology Plant Prot. 51(3-4):170-86.
 Mahmood, T., M. Arshad, M.I. Gill, H.T. Mahmood, M. Tahir and S. Hussain. 2003. Burewala Strain of
Cotton Leaf Curl Virus: A Threat to CLCuV Cotton Resistant Varieties. Asian J. Plant Sci., 2: 968-970.
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