Out look of Biofuels

1. Introduction

2. Types of Biofuels

• Bio-ethanol• Biodiesel

3. Classification of Biofuels

1st generation (Edible) 2nd generation*(Non-edible) 3rd generation (Algae)

4. Candidate Biofuel crops

5. JATROPHA*- A Promising Biofuel Crop

Genetic improvement of Jatropha

Designing Jatropha crop

6. Conclusion

Designer: A person who imagines, how something

could be made, draw plans accordingly, to design are

called Designer.

Designer crop: Possess specific characteristics, plan

before or during crop development e.g. Specific

protein, fat or starch quality, vitamin content,

elimination of anti-nutritional factor, color, taste of

grain/fruit; keeping quality or produce specific novel

biochemical or resistance to biotic and abiotic stress.

Introduction

Biofuel: Biofuel is produced from renewable biological

resources such as plant biomass and treated municipal and

industrial waste.

Solid, liquid or gaseous fuel consisting of Plant (tree

to grasses) its biomass (Wood chips to Seeds) – recently

Algae, Salicornia (marine weed) and Mangroves etc. also

being used to synthesize biofuel.

Why biofuels?

• Reduce dependence on fossil fuels

• Reduce reliance on foreign oil

• Biofuels reduce carbon dioxide emission up to 80%

• Biofuels produce 100% Less Sox/Nox than petroleum

• Biofuel reduce exhaust smoke emission and particulate matter by up to 75%; eliminate usual black cloud associated with fossil fuel

• Smell of biofuel exhaust is far more pleasant

Biofuel Advantages

Environmental Aspects:

√ Renewable

√ No sulfur (0.001%) – non toxic

√ Decreases GHG emissions (up to 70%)

√ Biodegradable

Socio-Economic Aspects:

√ Boosting profitability of Agriculture & livelihood

√ Increasing Energy Security

The most common biofuels

Bioalcohols : Biologically produced alcohols, mostcommonly ethanol and less commonly propanol andbutanol.

Ethanol fuel is the most common biofuel worldwide,particularly in Brazil, USA etc. Alcohol fuels areproduced by fermentation of sugars derived fromwheat, corn, sugar beets, sugar cane, molasses andany sugar or starch from which alcoholic beverages canbe made (like potato and fruit waste, etc.).

Biodiesel:Biodiesel is the most common biofuel in Europe, Brazil and USA. It is produced from oils/fats (Rapeseed, Mustard, Soybean and Flax) through trans-esterificationproducing fuel similar in composition to fossil/mineral diesel.

Feedstock's for biodiesel include animal fats and vegetable oils (Soybean, Rapeseed, Mustard, Flax, Sunflower, Palm, Hemp, Jatropha, Karanj, Mahua etc.

Bioethers

Bio ethers (referred as fuel ethers or fuel oxygenates) are cost-effective compounds act as octane rating enhancers.

Biogas

Biogas is produced by the process of anaerobic

digestion of organic material by anaerobes. It can be

produced either from biodegradable waste materials

or by the use of energy crops fed into anaerobic

digesters to supplement gas yields. The solid by-

product, digest ate, can be used as a biofuel or a

fertilizer.

Classification of Biofuels

Biofuels

3rd Generation2nd Generation(Non-edible Crops)

1st Generation(Edible Crops)

feedstock

corn

wheat Sugar crop Oil seed

Products

Bioethanol

Biodiesel Vegetable oil Bioether Solid biofuels

Feedstock• Non feed crop –

jatropha, castor, pongum, mahua

• Inediable waste product

• Saw dust

• Product• Cellulosic

• Biohydrogen

• biomethane

Feedstock• Algae-

- Botryococcus brauni

- Chlorella vulgeris

• Products• Algae fuel

First generation biofuels

• First-generation biofuels are biofuels made from sugar, starch, vegetable oil, or animal fats using conventional technology.

• The basic feedstock for the production of first generation biofuels are often seeds or grains such as wheat, which yields starch that is fermented into bioethanol, or sunflower seeds, which are pressed to yield vegetable oil for conversion into biodiesel

First generation biofuel controversies

• It is an amply proven fact that edible products or by-products should not be diverted for conversion into biofuels so as to address the hunger across and keep control over prices.

• Expanding biofuel production from edible sources will create imbalance in availability to humans and animals as well.

Increasing importance of Non-edible oils

1st Generation

Food Vs Fuel

Food Prices

Not sustainable in mediumterm

Scarcity of land & Water

2nd Generation

Non-edible

Cultivated in degraded lands and drought tolerance

High oil content

New source of income

Second generation biofuels

• Non-food species are considered as feedstock for second generation biofuels: Biodiesel from Jatropha, Karanja, Mahua etc. and Ethanol from cellulose (any non-food or food crops).

Advantage

• Eliminate competition for food and feed

• More efficient and environment friendly

• Less farmland is required

Useful by product

• Certain food product can be used, when no longer useful for consumption

• Hence, it is called as “advanced biofuels”

Increasing importance of cellulosic ethanol

• Abundant supply of biomass

• Use non-food crop and the waste from crops.

• 63 million ha waste land translates to about maximum potential of over 500 million t of cellulosic ethanol per annum (equivalent to 350 tons of petrol- Umaid et al.,2010)

• Cellulose used in commercial ethanol production

– Cell wall made up of………..

Third generation biofuels

• It is also called algae fuel.

• Algae are low –input , high-yield feedstock's to produce biofuels.

• Based on laboratory experiment

• 30 times more energy / acre > soybeans

• Strains- Botryococcus braunii, Chlorella vulgaris

Also Adds to Atmospheric CO2 reductions

Candidate Biofuel crops in India

1. Jatropha

2. Karanj

3. Mahua

4. Neem

Karanj

Constraints in Using Energy Crops other than

Jatropha

•Long Gestation Period

• No Breeding Efforts in Candidate energy

crops

• Selection of CPT’s and utilization –

very poor

• Only Pongamia involved in Network

project

• Mahua included two years back

• Neem – lot of initial work limited to

germplasm collection & conservation

Benefits of Jatropha

JATROPHAA Promosing Biofuel crop

Family: Euphorbiaceae 2n=22

Small shrub, woody plant, non-edible

Highly cross pollinated crop

Easy propagation

Drought tolerant and Insect resistant

Oil content - 28-45 %

In-consistant seed yield

Having similar properties of diesel.

It’s contains high amount of protein with a

well balanced amino acid composition.

Drawbacks of Jatropha

Still in semi-domesticated stage

Lack adequate genetic variation and non

availability of improved varieties, limit it’sprospects of being a successful energy crop.

Seeds and vegetative parts of Jatropha are toxic innature. Seeds contain toxic (phorbol esters) andantinutritional factors (trypsin inhibitor, phytate,lectin and curcin).

Methods Used for Genetic improvement of Jatropha

• Introduction

– No Introductions from Centre of Origin or diversity

– Managed and created variability in indigenous germplasm

• Selection

– Selected plants evaluated in provenance trials

– No provenance could be identified as such as variety

• Plants selected were involved in crosses to converge yield attributing

traits through

• Hybridization

– Intra and interspecific hybridization and backcrosses

• Mutation breeding

– No evidences of induced variability or traits in use except

– Spontaneous variation

• Biotechnological tool

– Genome mapped and Identified polymorphic types for hybridization

– Identified and isolated genes to develop transgenic

Status of Research

Global Scenario

• Latest Information:• Sun et al (2012) described genetic basis through QTL mapping using

J. curcas x J. integerrima crosses revealed: favoured alleles originate fromdiverse parents; transgressive segregation & complex genetic basis of traits.Advocated occurrence of linkage or pleiotrophy; suggested use of elitejatropha varieties as recurrent lines for transfer of favoured alleles andalleles for female flower/fruit number.

• The complete genome of Jatropha analyzed and sequenced (Sato et al., 2010). Lui et al. (2011) identified 18 QTLs for oil traits, 3 eQTLs of oleosinacid genes; which expresses with contribution rates (R2) 10% higher, controlling oleic acid, total oil to initiate MAB. Several studies on molecular analysis reported information on various aspects can’t be referred due to paucity of space

Status of Research

National Efforts• The hype period attracted world attention and huge plantations undertaken

with yield claims beyond potential, ignoring QPM. The MLT of provenances(DBT & NOVOD) identified elites types. Two centers JNKVV and TNAU (NOVODnetwork) succeeded in developing first generation hybrids and provedpotential through breeding.

• A decade of research witnessed papers on various aspects, but none onpractical breeding. Salomon & Ezradanam (2002) reported male:female ratio(29:1) and geitonogamy/xenogamy in Jatropha.

• Researches published on conventional methods (Swarup, 2006; and Gour,2007), and molecular analyses (Yadav et al; 2010 and Johnson et al; 2011) forgenetic improvement.

• Gour (2013) attempted hybridization involving J. curcas and J. integerrimagenerated variability followed by backcrossing to introgress desired traitsreduced male:female ratio; identified plants with high seed yield and parentallines nicking better in breeding.

Conventional Breeding

Widened genetic variability attempting Intra andInterspecific hybridization to address Key traits forDomestication and yield:

– Early flowering and fruiting types

– Reduce the unproductive branch length

– Enhancing Female to Male ratio

– Tailor plant with reduced height and productive branch length

– High Seed yield

– High Oil content

Conventional Jatropha Plant

Branches -22/22; 177 Capsules

55 Capsules in Single Branch

Excessive Branch Length Prior to Reproductive Phase

Dwarf and Conventional Plant Type

Different Morphotypes (Foliage Size and Density)

Plant with Normal LeavesPlant with Small Leaves

Small Vs. Normal Leaves Small Leaves Superimposed

Intraspecific Hybrids in Jatropha

Development of intraspecific hybrids and selection of recombinant lines using:

•J. curcas x J. curcas to exploit hybrid vigour expressed n F1

– crosses revealed vegetative and reproductive vigour

– Bi-clonal plantation of two such plants will be done to realize potential in terms of high oil and seed yield

•J. curcas x J. curcas with unique quality traits viz.

– Non toxicity to utilize oil and oilcake &

– Non pigmented foliage, Dark green stem, Yellow stem, small leaves Flower colour for variety identification and IPP

Selection of desired recombinant in segregation population and Identification of superior genotypes-CPT’s will be multiplied clonally

Interspecific Hybrids in Jatropha

• Development of interspecific hybrids and selection of desired recombinant lines using:

• J. curcas x J. integerimma crosses and backcross with J. curcas

Desired recombinants are under rigorous evaluation forIdentification of Superior Genotypes

Inter-specific Hybridization

X

P1 P2

F1

F1’s

Mutant

Variability Generated Through Intra and Interspecific Hybridization

Designing Jatropha Crop

Early flowering and fruiting types - Achieved

Reduce the unproductive branch length - Obtained

Enhancing Female to Male ratio - Achieved

Tailor plant with reduced height & productive branch length - AchievedDwarfing for Effective Management and High Density

plantation

High Seed yield – Achieved targeting still higher

High Oil content - Achieved

Non toxic – On way

Synchronous flowering/fruitingSynchrony within bunch - Achieved

Indeterminate growth – Challenge to achieve

Genetic Efforts and OutcomeTailored Designer Plant

JJH H 1-5

Breeding Non-Toxic Lines

Phorbol Content

•The phorbol estimation through HPTLC/HPLC incrosses between nontoxic (shy bearer) and toxiclines is being done.

• The estimates of phorbol content depicted nodetection (NT 61-3) or very low (NT 52-2) ascompared to standard.

•Plants with Lower/no phorbol content isolated withhigher fruit bearing are being analyzed.

Estimation of Phorbol ester

HPLC Standard: phorbol-12-myristate 13-acetate

Peak results:

Cont..

Phorbol ester Sample peak : Jc 11 x NT (1)

Peak results:

Analysis of Plants for Non-Toxicity by HPTLC

34-6 21-8 G3-3 9-1(old) H1-5 61-3

(seed

s)

61-3

(leave

s)

H2-1 G4-7 G2-7 G2-3 G1-8 H1-6 H9-9 Dwarf PMA

1510.8 1202.

1

1069.9 606.5

VIII

931.

6 II

418.0 1840.7 761.2

V

823.4

III

810.7

IV

3271.1 768.

8 VI

957.6

I

709.

2 VII

1096.8 985.9

JC15x

CF1(1)

JC4x

BCF1

(1)

552-3 x

34-6

NDCJC1

x 16

9-

1(old)

x 61-

3

JC13x

NT 6

JC13x

NT 6

17-21 x

61-3

52-5 x

61-3

61-3 x

52-2

52-3 x

34-6

61-3 x

?

9-1(old)

x 61-3

52-2 x

?

Sponta

neous

Stand

ard

Genetic Enhancement of Yield and

Beyond to maximize Production

Using BA

JJH 34-6 with 54 Female

Flowers

JJH 34-6: 54 Female Flowers and >36 Developing Capsules Variety Plus BA Effect

1

23

4

5

6

7

89

1011

1213

14

16

19

18

17

15

20

30

29

28

27

26

2524

23

22

21

31

32

33

34

35

Conclusion

The Designer plant with availablevariability and conventional breeding approachand intensive selection have paved way totailor Jatropha plant with reduced heightsuitable for high density plantation combiningearliness, high female to male flower ratio,high seed and oil yield per unit time and spacewith easy management.

There still exist scope for furtherdevelopment of improved and non-toxicvarieties using frontier technologies to fosterdevelopment of new improved varieties withnew ideas and innovations.

Thank ‘U’

JNKVV Few Step Ahead ……… in Transforming and Genetically Designing Jatropha for Domestication