Captain Green Reducing the global warming gases and increasing - - PowerPoint PPT Presentation

The Team: E. Varma, S.Tejpal, S. Jha, P. Rana, P. Sharma, N. Aggarwal, V. Manohar, P. Sharma, M. Sharda Professors:

• Dr. Antresh Kumar
• Dr. Saurabh Bansal
• Dr. R.S Chauhan

Captain Green

Reducing the global warming gases and increasing fertility of paddy fields

J U I T India

CO2, 70% N 2O, 20% CH4, 9% Others, 1%

Global Green House Gases Em ission

CO2 N2O CH4 Others IPCC Fourth Assessment Report, 2008

Energy Supply- Main contributor

Effects (concentration – 1 to 8 %) Drowsiness, Increased blood pressure, Dizziness, shortness of breath, panic attacks, headaches.

Adopted from U.S. Environmental Protection Agency

CO2 emissions from industrial processes CO2 emissions from land use change

Source: www.oup.co.uk/vsi

Agricultural Soil - Main Contributor of N2O

Natural Nitrous oxide sources ~ 10 million tonnes each year Man-made Nitrous oxide sources ~ 8 million tonnes each year. Effects (>100ppm) Asphyxiation, Kidney damage, headaches

Adopted from GHG online

Rice Cultivation- Main contributor of CH4 emission

29% 8% 6% 4% 4% 3% 3% 11% 18% 14% Rice cultivation Biomass burning Landfills Sewage treatment Animal waste Termites CH4 hydrates and ocean Wetlands Coal & miming/ natural gas Enteric fermentation

Effects (Dangerous at 5-15%concentration in air) Dizziness, headaches, nausea, drowsiness and unconsciousness

Adopted from National Aeronautics & Space Administration

FAO Estimates 2010

2010 estimates FAO estimates 2010

FAO Estimates 2010 International Grains Council estimates, 2012

Rice production in India increased from 44.69 million tons (2001) to 95.32 million tons (2011).

Sharad Pawar, Agriculture Minister, 2011

Rice is a high energy calorie food.

• Major part of rice – carbohydrate (starch) - 72-75%.
• Protein (glutelin) content of rice is 7%
• Nutritive value of rice protein (biological value = 80)
• Rice contains most of the minerals ( 4% phosphorus).
• Rice also contains some enzymes.
• India has become one of the top five countries with rice exports (2011).
• Employment of a large number of people.
• Products of the rice plant are used for a number of different purposes, such as
• fuel, thatching, industrial starch, and artwork.

0% 10% 20% 30% 40% 50% 60% 70% 80% Methane Nitrous oxide Carbon dioxide Contribution to global warming Contribution by paddy fields

H% GHG emission

Adopted from GHG online & U.S. Environmental Protection Agency

Methanogens Bacillus Sp. R h i z

• b

i u m s p . Cyanobacteria sp. Methylococcus capsulatus

Thermo Tolerancy Denitrifying gene present MMO encoding gene. Facultative Anaerobe

Methanotrophic bacteria – R.S. Hanson and T.E. Hanson, 1996,60(2)

Paddy and the microbes

Captain Green: To Do List – Convert CH4 into

• CO2 at a faster rate.

Convert N2O coming

• from the nitrogen

fertilizers into NO3. Constitutively show

• it’s skills to be really

called a captain and show no competition with the natural flora.

MxaF NosZ NifA SacB O U R C O N S T R U C T

MxaF

Present in Methanotrophs

• Located upstream of genes of
• Methanol Dehydrogenase

Methanol Inducible Promoter

• Create a diversion of flux
• Leading to faster degradation of
• methane.

MxaF CH3OH

N

• s

Z

Present in Denitrifying

• Bacteria

Responsible for

• conversion of nitrous
• xide into free nitrogen.

N2O N2

NifA

Transcriptional

• Activator for Nif genes

Nif genes are a cluster

• f genes responsible for

conversion of free nitrogen into nitrate i.e. nitrogen fixation.

S a c B

Used as a Negative selectable

• marker

Codes for levansucrase

• Levansucrase is a transfructosylase
• catalyzing sucrose hydrolysis and

levan synthesis The gram negative microbe cannot

• survive in the presence of sucrose

CH4

CH3OH

MxaF nosZ nifA (Promoter)

N2O N2 NO3 inducer

Methylococcus Capsulatus

OUR APPROACH

NH2OH

T H E EX P E R I M E

• 1. Design of primers & Optimization of PCR

reactions

MxaF NosZ SacB NifA

SpeI SpeI AceI AceI NdeI NdeI AscI AscI

MxaF NosZ NifA

Y X Y Z Z Y X Ligation Nos Z NifA Ligation of Gene Complete Fragment

SacB

Genomic DNA

THE EXPERIMENT

The Detection Systems

T H E EX P E R I M E

Gas Chromatography for detection of methane and nitrous oxide. Flame Ionization Detector - methane

• Thermal Conductivity Detector - nitrous oxide.

References

Genomic Insights into Methanotrophy: The Complete Genome Sequence of Methylococcus

• capsulatus (Bath) - Naomi Ward et.al

Over-production and Characterization of the nifA Gene Product of Klebsiella pneumoniae -

• the Transcriptional Activator of nif Gene Expression By R. T U L I t AND M. J . MERRICK

Global Methane Emissions From Wetlands, Rice Paddies, and Lakes* - EOS, vol 90 no. 5, 3 feb

• 2009

Methanotrophic bacteria - R S Hanson and T E Hanson- Microbiol. Mol. Biol. Rev. 1996,

• 60(2):439.

Methane and Nitrous Oxide Emissions from a Rice Field in Relation to Soil Redox and

• Microbiological Processes - A. X. Hou, G. X. Chen, Z. P. Wang, O. Van Cleemput, and W. H.

Patrick, Jr.*

Suggestions & Comments

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