Biotechnologydriven sustainable agriculture Dr SHAHID MANSOOR SI HEC

Biotechnology-driven sustainable agriculture Dr. SHAHID MANSOOR, SI HEC Distinguished National Professor DIRECTOR National Institute for Biotechnology and Genetic Engineering (NIBGE), Faisalabad, Pakistan 1

Sustainable agriculture; Challenges and opportunities Challenges • Population growth and food security • Yield stagnation and increasing cost of production • Climate change and environmental degradation • Water availability and quality • Salinity and water logging • Land availability • Pest and diseases Opportunities Major progress in understanding of genetics, genomics and genetic engineering Challenges bring opportunities 2

NIBGE Ø An affiliate centre of ICGEB, Trieste, Italy Ø Center of Excellence by Min. of Science & Technology Ø National library of Biological Sciences Ø Affiliation with Quaid-i-Azam University and Pakistan Institute for Engineering and Applied Sciences (PIEAS) M. Phil & Ph. D Biotechnology Degree Ø ISO 9001 -2008 Certified

Partners; PAEC Agri. /Biotech Centers/other strategic organizations

Crop Improvement strategies Conventional Green revolution Conventional breeding Mutation breeding High yielding varieties responsive to fertilizers and pesticides Plant Breeding Gene Pool, Limited Biotechnological Gene revolution Tissue Culture Genetic Engineering Marker-assisted selection Crops with high yield and efficiency Genetic Engineering Gene Pool, Unlimited

Genomics; a silent revolution Major improvements in sequencing technologies All major crops/livestock have either been sequenced or will be sequenced in the next few year • • DNA based marker technologies Genotyping by sequencing (GBS) Automated phenotyping Bioinformatics; Our ability to handle genomic data Huge potential for crop improvement, Disease diagnosis and environment

How technologies can help in food security and sustainable agriculture in irrigated and non-irrigated areas? • Genomics and understanding of QTLs and genes affecting productivity • Combination of genetic selection and genetic engineering in evolution of superior varieties • Technologies to reduce cost of production/yield losses • Sustainable improvement of soil health and underground water resources • Selection of high performing livestock through genomics • Quality/safety enhancement of products • Mechanism of transfer of technologies

Success stories from Pakistan Genetic engineering Cotton Wheat Sugarcane Potato Genomic selection Rice Wheat

NIBGE efforts in developing Bt cotton varieties 2014 Upcoming lines New IR-lines With better fiber 2007 IR-NIBGE-3701 • National variety, approved in 2010 & 11 • 25%, 18% area in 2010, 2011, respectively • 1 -1. 2 million bale advantage Ti m e lin e Total Approved cotton varieties IR-NIBGE-5 2013 1. NIBGE-2— 2006 IR-NIBGE-6 2. IR-NIBGE-1524— 2010 • Completed on year in NCBT 3. IR-NIBGE-3701— 2010 2011 IR-NIBGE-4 • Tolerance to lodging rotenning 4. IR-NIBGE-901— 2011 5. IR-NIBGE-3— 2012 • Approved in 2012 2010 IR-NIBGE-3 • Popular for spring cultivation 6. NIBGE-115— 2012 7. NN-3— 2013 2006 2005 2002 IR-NIBGE-1524 Improved IR-NIBGE-901 IR-FH-901, IR-448 & IR-443 • National variety approved in 2010 and 2011 • Drought prone area (2%) • Approved for Sindh in 2011 • 20% (2009 & 2010) • 25% in 2011

Summary of the Impact Sindh (A) Million bales Million US$ Punjab (B) Million bales Million US$ IR-NIBGE-901 2. 14 858. 96 0. 235 75. 83 IR-NIBGE-1524 0. 09 31. 91 0. 822 265. 02 IR-NIBGE-3701 0. 37 138. 98 4. 714 2115. 63 Total 2. 60 1029. 84 5. 771 2456. 48 Million bales Million US$ 8. 37 3486 Grand Total (A+B) = US$= Rs= 8. 37 Million bales 3486 Million or ~345 Billion

Limiting factors and cotton genetic engineering Virus resistance Multiple gene construct (G 5, Gro. EL, artificial zinc finger Insect and herbicide resistance Cry 1 Ac+Cry 2 ab+EPSPS Sucking pests Lectin and Hvt under phloem-specific promoters RNAi based resistance against whitefly Abiotic stress tolerance Three gene construct for drought and salt stress tolerance Fiber improvement Three gene construct for fiber length improvement

Rice Major constraints include: Abiotic Biotic Drought Submergence Salinity Phosphorus deficiency Diseases Bacterial leaf blight Bacterial leaf blast High priority at NIBGE • Provision of services to rice exporters • Understanding genes that contribute to yield • Improved crop varieties through DNA markers

Advance Line 1 Advance Line 2 BLB resistant Basmati advanced lines

Fine Rice NUYT Trial-2013 Paddy Yield (Kg/ha) Designation 1 2 3 4 5 6 7 BR-1 PK 8431 -2 -4 BR-18 R-456 BR-23 PK 8647 -11 -1 -1 PK 8892 -4 -2 -1 -1 8 9 PK 8685 -5 -1 -1 NIAB-1175 Trial Locations RRI NARC Dokri 3066. 7 de 3500. 0 d 6073. 3 ab 6395. 0 a 2793. 3 ef 4083. 0 cd 5260. 0 abc 6000. 0 a 1586. 7 f 4000. 0 cd 6280. 0 a 4583. 0 bc 4306. 7 cd 4500. 0 bc Sujawal 6218. 0 ab 4630. 0 cd 6633. 0 a 4519. 0 d 5700. 0 b 3952. 0 ef 5078. 0 c Average NIAB Faisalabad 2832. 1 cd 2774. 6 de 2545. 9 f 3139. 5 ab 3315. 2 a 2552. 8 ef 4683. 2 a 4554. 0 ab 4491. 7 ab 4373. 6 bc 4188. 0 cd 4165. 0 cd 4058. 3 d 4180. 0 c 4066. 7 c 3642. 0 g 3813. 0 f 3879. 7 d 3008. 3 bc 3547. 0 e 3292. 6 a 4046. 7 d 4037. 9 de 2770. 0 g 3925. 0 e 4411. 6 c 3794. 2 ef BASMATI-515 CHECK 4100. 0 cde 4500. 0 bc 4363. 0 de 2366. 7 h 3533. 0 h 4765. 4 b 2798. 3 cd SUPER BASMATI Check Location means 3076. 7 fg 4450. 0 c 3690. 9 de 2827. 7 cd 10 PK 8667 -8 -5 -1 -1 5433. 3 abc 4583. 0 bc 3599. 0 f 4700. 0 bc 4333. 0 4512. 0 d bcd 4180. 0 cde 5000. 0 b 3715. 0 f RRI ARF Tando Jam Gujranwala KSK 6566. 7 a 4921. 3 b 5677. 6 a 3080. 0 fg 4529. 0 c 4394. 9 c 5590. 0 b 4221. 0 d 5575. 2 a 3693. 3 d 3704. 1 g 4258. 7 c 5936. 7 b 3988. 0 e 4965. 1 b 3580. 0 de 3675. 0 g 3768. 7 d 3290. 0 ef 5021. 0 a 3659. 4 de 3273. 3 de 3583. 0 d 4580. 0 cd 2557. 4 ef 3775. 2 f 3640. 4 f 4254. 4 cd 4588. 5 ab 4792. 1 a Means with the same case letter do not differ significantly at p<0. 05. 4016. 4 d 4118. 6 cd 4382. 9 c 2902. 0 e

BLB resistant Super Basmati developed through genomic approach showing resistance in field

Wheat Biotechnology at NIBGE • Drought tolerance Genes • AVP 1/AVP 1 -D • HVA 1 • DREB 1 A Procedure for Transformation of wheat Percent increase 25 -30 % 13 % Transgenic drought tolerant wheat AVP 1 and AVP 1 D genes 5 -10 % • Salinity tolerance • At. NHX 1 • HVA 1 15 -30 % 20 % Testing of drought and salt tolerant wheat in the field Transgenic salt tolerant wheat At. NHX 1 and HVA 1 genes

Wheat biotechnology Understanding of genes contributing to yield • pyramiding of genes through genomic selection • genetic engineering Order of Priority 1. Phosphorus use efficiency 2. Heat tolerance 3. Drought tolerance 4. Rust resistance 5. Salt tolerance 6. Herbicide tolerance Marker-Assisted Breeding Order of Priority 1. Rust resistance 2. Drought tolerance 3. Salt tolerance

Genomics/ DNA markers for wheat improvements 2010 -13 Strain name Maturity(DH*) Disease Height (cm) Color Spike (spikelet) Yield Maunds/Acre) NN-Gandam-1 85 -87 DH Dominan tly Free 90 -95 Light green Compact (23) 65 (normal sowing) 45 (late planting) NN-Gandam-2 82 -85 DH Free 95 -100 Dark green Relatively loose (21) 67 (normal) 37 (late) Sehar 90 -95 DH Susceptib le to LR 96 -103 Green Compact (19) 58 (normal) 33 (late) • • • DH= Days to heading Normal= planted on Nov 10 Late= planted on 25 Dec Wheat: Achieving food security through developing • High yielding varieties • Drought tolerant varieties • Heat tolerant varieties • Early maturing varieties • Resistant to rust

Vegetatively grown crops; Transgenic sugarcane and potato for biotic and abiotic stresses Sugarcane; major thrust Insect resistance/herbicide tolerance Drought / salt / Frost tolerance Screening of transgenic sugarcane lines under field condition (two years trial) • Drought and frost tolerant (20% water saving) • Salinity tolerant Future thrust • Sugarcane with multiple transgene • Addition of legumes such as soybean and mungbean (intercropping)

Drought Tolerant Sugarcane expressing AVP 1 gene At 100%Irrigation Sr. # At 80%Irrigation At 60%Irrigation Variety Cane Yield (t/ha) Control Transgenic %Increase 1 S 2003 US-114 130 132 1. 54% 2 HSF-240 122 125 2. 46% 3 CPF-246 131 135 3. 05% 4 CSSG-668 92 104 13% Sr. # Variety Cane Yield (t/ha) Control Transgenic %Increase 1 S 2003 US-114 110 124 12. 7% 2 HSF-240 109 118 8. 26% 3 CPF-246 119 127 6. 72% 4 CSSG-668 88 98 11. 4% Sr. # Variety Cane Yield (t/ha) Control Transgenic %Increase 1 S 2003 US-114 93 108 16. 1% 2 HSF-240 89 102 14. 6% 3 CPF-246 98 110 12. 2% 4 CSSG-668 73 85 16. 4%

Future directions • Input use efficiency • Diversity of genes and promoters • Multiple genes • Integration at defined loci • Chloroplast biotechnology Capacity enhancement in biosafety assessment • Regulatory framework • Capacity enhancement in biosafety evaluation

Improving soil health and sustainability • Reduction in use of chemical fertilizers • Enhancement of organic matter • Minimize environmental foot print by degrading pollutants (insecticides/herbicides)

Biodegradtion of Pesticides (Plantmicrobe interactions) Degradation of chlorpyrifos in rye-grass vegetated soil inoculated with Mesorhizobium Sp. HN 3 yfp Control (No CP) Biodegradation of bispyribac. Na in wheat vegetated soil (pot experiment containing 2. 5 Kg soil) EBD 50 mg/Kg CP + culture

Range of Products

Bio. Power Production Technology transfer through ATCOP to ICL Future: Next generation Biofertilizer with value additions, e. g. , biopesticides, growth promoting hormones, P- solublization, new carrier materials and organic matter EBD Field testing of Bio. Power

IMPROVEMENT OF LIVESTOCK & POULTRY PRODUCTIVITY Livestock and poultry; health and production in Pakistan can be improved by Ø Establishing early and sensitive diagnosis of infectious diseases Ø Development of effective animal vaccines. Ø Genetic improvement of livestock breeds for milk and meat (i. e. , whole genome sequencing of sahiwal cattle). Ø Improving digestibility of fibrous animal feed (enzymes, probiotics and prebiotics).

Diagnosis of infectious diseases Pioneer research group in the country which has established PCR based diagnostic facilities for • • Rinderpest Peste des Petits Ruminants Foot & Mouth Disease Canine Distemper Infectious Bursal Disease New Castle Disease Avian Influenza Brucellosis

Use of genotyping by sequencing (GBS) for selection of high performing cattle NIBGE, RCCS and CAAS-ILRI Joint Laboratory on Livestock and Forage Genetic Resources Institute of Animal Science (IAS), Chinese Academy of Agricultural Sciences (CAAS) Objectives • Selection of proven bulls • Selection of high performing cows • Genetic improvement through crossing and genomic selection

Genetic Improvement of Livestock Breeds Whole genome sequencing of Sahiwal cattle in collaboration with Jamil-ur -Rehman Institute of Genomics, University of Karachi, Karachi and Research Center for Conservation of Sahiwal Cattle (RCCSC) Jhang

Improving Digestibility of Fibrous Animal Feed Intra institute collaboration with Probiotic and Food Safety group and Industrial Biotechnology Division for production of enzymes, probiotics & prebiotics (Dr Farooq Latif, Dr Hamid, Dr Arslan, Dr Munir)

Wheat enhancement project (IDB) Saudi Arabia (KAUST) Pakistan (NIBGE) UAE (ICBA) Egypt (Damanhour University) Objectives • Yield increase by genomic selection and genetic engineering • Germplasm and elite lines with enhanced nutritional value • biotic stress tolerance (drought, salt, heat and frost) • Enhancing biotic stress tolerance against rust diseases and other fungal diseases Progress Ten clones for heat, drought and salt tolerance USD 20, 000 to be provided by KAUST

Use of genomic resources for optimizing the input use in cotton Collaborative countries: Uzbekistan, Sudan, Nigeria, Egypt, Turkey & Pakistan Vaccines for animal and human diseases Iran and Pakistan

Take home message Major investment in genomics and genetic engineering for food security and sustainability Crop varieties High yielding, water efficient disease resistant, high efficiency for fertilizers Sustainability Reducing cost of production by Biofertilizers Reducing environmental impact by degradation of agrochemicals Livestock and poultry Breeds for milk and meat Better health by vaccines, probiotics Enhancement of feed conversion

Thanks