Brassinosteroids plant harmones

2. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-2 Advertising  Paid, nonpersonal communication through various media by business firms, not-for- profit organizations, and individuals who are identified in the advertising message and who hope to inform or persuade members of a particular audience Seminar on Brassinosteroids- New Class of Plant Hormone Presented By Sonali L. Bagade Submitted To DEPARTMENT OF AGRICULTURAL BOTANY POST GRADUATE INSTITUTE DR. PANJABRAO DESHMUKH KRISHI VIDYAPEETH, AKOLA – (M.S.)

3. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-3 Introduction Steroid-: These are the compounds which are widely distributed in animals & plantshave 1,2- cyclopenteno-perhrydro- phenantherene nucleus there. structure 1 ) It plays a very imp role as essential hormones in plants as well as animals. 2 ) Plants produce numerous steroids& sterols, some of which recognized as hormones in animals (Geuns,1978; Jones and Roddick 1988). Brassinosteroids-: Brassinosteroids are a new group of plant growth substance with significant growth promoting activity (Grove et al. 1979) . Pollen – Rape seed( Brassica nupus L ), for determine it’s structure. the biologically active molecule was a steroids that calls Brassins. Since there discovery over 70 BR have been isolated from plants . It is biosynthesized from compesterol. . Arabidopsis, pea & Tomato plans are used . BRs have been shown to be involved in numerous plant processes:

4. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-4 Commercially available Brassinosteroids Trade name country Recommended crops Biobras-6 Cuba potato, sugarcane & onion Bountee India Tea Brassinosteroids 1105 USA Soybean Combine India Grapes DAA-6 Cuba Potato ,tomato & sugarcane Double India All major crops Ts 303 Japan Rice, potato, wheat, beet Yaponiya Belarus Barley

5. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-5 Immediately after the discovery of Brassinosteroids the application of this substance for improving crop productivity of diff. vegetables like radish ,lettuce, bushbean and paper was started. Since the publication of this article “New chemicals promise larger crops.”(Maugh1982) to “Twenty years of Brassinosteroids.” (Kripach et al). However ,promoted by the success stories ,several agrochemical industries in Japan USA, Belarus, Cuba and India recently started commercial production of BRs, at affordable level. “The shadow of flop story- triacontanol also put a ride on the commercial production. Uses- Improve resistance against envt. Stresses – 1. water stress 2. Salinity stress 3.Low temp stress 4. High temp stress (Rao et al 2002) Ability to confer resistance against abiotic stress added advt. for uses in agril. For improving crop Agricultural uses of Brassinosteroids

6. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-6 Brassinosteroids-Potential to improve crop productivity Crops Frit crops Grapes Orange Citrus Strawberries Watermelon Banana Compound Epibrassinoloide Brassinolide Epibrassinoloide Brassinolide Brassinosteroids Epibrassinoloide Biobras-16 Response Yield Reduced abscission of young flowers, early maturation Growth & yield Retardation of abscission increased fruit lets Retardation of abscission increased fruit lets Yield In vitro multiplication Reference Ikekawa and zhao 1991 Xu et al. 1994 Takshashi et al. 1995 Iwahari et al.1990 Franco et al.2002 Pipattanawong et al.1996 Ikekawa and zhao 1991 Rodriguez et al 1998

7. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-7 Brassinosteroids-Potential to improve crop productivity Crops Veg.crops Lettuce Bush bean Paper Soybean Garlic Potato Yam Spinach onion Tomato Lettuce Tomato Compound Brassinolide Brassinolide Epibrassinoloide Homobrassinoloide Brassinoloide Epibrassinoloide Biobras-6 Brassinosteroids DA-6 Epibrassinoide DA-6 Biobras-16 Biobras-6 Biobras-16 Homobrassinoide Response Growth & yield Yield Yield Growth & yield Yield Tuber Yield Tuber yield Growth & yield Bulb yield Yield Yield Accelerating of ripening Reference Meudt et al.1983 Meudt et al.1983 Meudt et al.1983 Nakaseko & Yoshida 1989 Nunez et al.1994 Nunez et al.1995 Labrada 1997 Liang et al.1998 Nunez et al.1998 Nunez et al.2000 Augustin 2001 Vardhini & Rao 2002

8. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-8 Brassinosteroids-Potential to improve crop productivity Crops cereals Wheat Rice Maiz Barley Compound Brassinolide Epibrassinoloide Homobrassinoide Brassinolide Epibrassinoloide Homobrassinoide Biobras-6 Brassinolide Epibrassinoide Biobras-16 Epibrassinoloide Response Yield Yield Growth & yield Grain Yield Yield Yield Yield Growth & yield Growth & yield Growth & yield Reference Braun and Wild 1984 Ramraj et al. 1997 Sairam 1994 Yokota and Takashashi 1986 Ramraj et al. 1997 Franco et al.2002 Takshashi et al. 1995 Ikekawa and zhao 1991 Almenares et al 1999 Lapa et al 1998 Prusakova et al.1995

9. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-9 Brassinosteroids-Potential to improve crop productivity Crops Oil Seed Groundnut Mustard Lens Other crops Tobacco Sugercane Sugerbeat Cotton Chickpea Compound Brassinolide Epibrassinoloide & Homobrassinoide Homobrassinoide Homobrassinoide Brassinolide Biobras-6 Biobras-16 Brassinosteroids Response Growth & yield Nodulation & nitrogenase activity Yield Yield Leaf yield Cane propagation Yield Yield Proteins and yield Reference Vardhini & Rao 2002 Hayath et al 2000 ,2001 Hayath and Ahmed 2003 Yokota and Takashashi 1986 De laFa-CF et al. 1998 Schilling et al 1991 Ramraj et al. 1997 Ramos 1997

10. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-10 Effect of BRs under abiotic & biotic stresses Stress plant Abiotic stress Drought/water Sugar beat stress Wheat Chick pea Sorghum Maiz Mustard Salinity stress Sugar cane Rice Wheat Compound Homobrassinoide Homobrassinoide Epibrassinoloide Epibrassinoide Epibrassinoloide Brassinolide Epibrassinoloide Homobrassinoide Brassinolide Brassinosteroids DAA-6 Brassinosteroids Epibrassinoloide Effect increased growth & Yield Increased yield Enhanced growth & yield Enhanced growth increased growth Alleviation of stress impact on seedling growth Increased tolerance Increased oil yield Enhanced growth & yield Improve tolerance Growth recovery Reference Schilling et al.1991 Sairam 1994.1996 Prusakova et al.2000 Singh et al.1993 Xu et al 1994 Vardhini & Rao 2003 Li and standen 1998 Kumawat et al 1997 Gonzalez & Gianza1997 Takeuchi 1992 Benrukova 1998

11. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-11 Effect of Brassinosteroids under abiotic & biotic stresses Stress plant High temp. Wheat Rape seed Chilling stress tomato Maize Tomato Rice Wheat Heavy metal Barley stress Compound Brassinosteroids Epibrassinoloide Epibrassinoide Epibrassinoloide Brassinolide Epibrassinoloide Brassinosteroids Brassinosteroids Effect Enhanced photosynthetic rate & nitrate reeducates activity Increased stress tolerance Increased stress tolerance Increased stress tolerance Alleviation of stress impact on seedling growth Alleviation of stress and increased membrane stability Enhanced Reference Kuleva et al.1991 Dhaubhadel et al 1999 Dhaubhadel et al 1999 He et al 1991 Kamuro & Takasuto 1999 Xu et al 1994 Vardhini & Rao 2003 Wang and Zang 1993 Li and standen 1998 Kumawat et al 1997 Khripach et al 1999

12. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-12 Effect of Brassinosteroids under abiotic & biotic stresses Stress plant Herbicides. Rice Biotic stress Tobacco Potato Compound Brassinosteroids Brassinosteroids containing leaf extracts Epibrassinoloide Effect Herbicides safener Increased stress tolerance Enhanced resistance against fungal pathogens Enhanced resistance against mixed fungal pathogens Reference Kim et al.1993 Roth et al.2000 Volynets et al 1997

13. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-13 Unlimited potential of Brassinosteroids- 1. Inhibitors of nucleic acid synthesis (actinomycins) and protein synthesis (cyclohexamide) were found to set aside the epibrassinolide- stimulated growth in mungbean epicotyls .(Wu and Zhou 1993) 2. Epibrassinolide stimulated growth of Chinese cabbage mesophyll protoplast was associated with enhanced protein levels .(Nakajima et al. 1996) 3. a) BRI 1 is a leucine rich repeat receptor kinase (LRRRK) known to recognize protein ligands .( Li and Chory 1997). b) The growth of brassinosteroid deficient dwarf mutant lkb of Pisum sativum was restored with the exogenous application of brassinolide .(Nomura et al. 1997) c) Subsequently castasterone, another steroidal substance was isolated from the insect galls of chestnut (Castanea crenata) . As the 1st steroidal plant growth regulator was isolated from Brassica napus, a generic name ‘brassinosteroids’ has been given to this new group of phytohormones. Brassinosteroids are polyhydroxysteroids. They have a common α- cholestane skeleton and their structural varieties come from the kind and the orientation in the A/B rings and side chain . (Yokou,1997)

14. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-14 Unlimited potential of Brassinosteroids- 5. a) The studies conducted with brassinosteroid biosynthetic mutants provided compelling evidence for the essentiality of this new group of growth regulators for the normal growth and development. The growth of BR-deficient dwarf mutant of Arabidopsis such as cpd, dwf I rescued by brassinolide . (Bishop et al 1999) b. Brassinosteroids are considered as plant hormones with pleiotropic effects as they influence wide array of developmental processes such as growth, seed germination, rhizogenesis, flowering, senescence, abscission, and maturation . ( Sasse 1999) 6. Inhibitor studies employing brassinozole, a specific inhibitor if brassinolide in plants, further strengthened the view that brassinosteroids are essential for growth and development in plants. (Assami et al. 2000) 7. The dwarf tomato mutant dumpy (dpy) , a dwarf with abnormal leaf phenotype (curled with dark ruggose leaves) with the suppression of axillary shoots was found rescued to wild type with the foliar application of brassinosteroids. ( Bishop and Yokota 2001 )

15. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-15 Unlimited potential of Brassinosteroids- 8. a) Brassinosteroids satisfy all the prerequisites to be considered as plant hormones i.e. natural in occurrence, mobility and activity in extremely low concentrations . (Rao et al 2002 ) b) Rinrei, is a dwarf mutant of faba bean (Vicia faba) characterized by dark green leaves and reduced plant height, contributed by a single recessive gene mutant and the gene was tentatively identified as brassinosteroid defient dwarf (bdd). (Fukuta et al. 2002) c) Brassinolide, 24-epibrassinolide and 28-homobrassinolide are the three active brassinosteroids, being widely used in physiological studies. Brassinosteroids have been reported from 60 species which includes 51 angiosperms ,12 monocots and 39 dicots , 6 gymnosperms, 1 pteridophyte ( Equisetum arvense strobillus ), 1 bryophyte ( Marchantia polymorpha ) and 1 alga ( Hydrodictyon reticulatum ) . ( Rao et al. 2002; Bajguz and Tretyn 2003 ) 9. A) 65 brassinosteroids and 5 sugar fatty acid conjugates have

16. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-16 Distribution of BRs in Plant kingdom Plant species Plant parts Brassinosteroids Monocot- Oryza sativa Shoot 1, 4,,25 Triticum asestivum seed 11, 28, 29 Zea maiz grains 4, 8,9,10,11 Dicot- Arabidopsis thaliana shoot 4,1 Bracssica napas pollen 1,4 Dolchios lablab seed 1,4,11,24-26,30,31 pisium sativum seed 1,4, 8,11,18 Gymnosperm- pinus thunbergji pollen 4,8 picea sitchensis shoot 4,8 Algae- hydrodictyon reticulatum collony

17. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-17 Arabidopsis phenotype Figure 8. Phenotype of Arabidopsis antiTRIP-1 transgenic lines. a) Two- month-old α-TRIP-A (left) vs. wildtype (right), showing the extreme dwarfism and altered leaf morphology of the antisense line, similar to BR-deficient and – insensitive mutants. b) Six-month-old α- TRIP-D showing delayed senescence and bushy growth habit similar to the bri1 mutant. c) Two-month-old α-TRIP-C1 showing the abnormally high number of rosette leaves and the initiation of multiple floral meristems before the elongation of the inflorescence. d) The arrow shows the initiation of a new rosette of leaves and multiple inflorescences at the terminal point of a primary inflorescence in α-TRIP- C1. e) Line α-TRIP-C1 initiated an abnormally high number of floral buds

18. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-18 Phenotype of the bri1 mutant Figure 6. Phenotype of the bri1 mutant. Panel A shows 2-month-old mutant (left) and wild type (right) plants grown in 50 ml centrifuge tubes in a 23 C growth chamber (16 hr light / 8 hr dark). Panels B and shows close-up view of a 2-month-old bri1 mutant plant. Panel C shows the same plant after 4 months. All bars represent 1 cm. Adapted

19. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-19 Dwarf Phenotype of bri1 Mutants in Different Plant Species Figure 3. (A) Six-week-old wild-type and bri1- 1 mutant Arabidopsis plants. (B) Twelve-day-old wild-type and lka mutant pea plants. (C) Seven-week-old wild-type and cu-3 mutant tomato plants. WT, wild type. Bars in (A), (B), and enlarged inset photograph of cu-3 in (C), 5 cm; bar in (C), 15 cm. Plant Cell. 2002; 14(Supplement): s97–s110. doi: 10.1105/tpc.001461. Copyright © 2002, American Society of Plant Biologists

20. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-20 BRs on salinity induced growth inhibition of groundnut seedlings BRs, 24- epibrassinosteroide & 28-homobrassinosteroides were produced from Beak consultants Inc:Brampton Ontarno, Canada .Seeds of groundnut (Arachis hypogaea L.Var ICGS44) were obtained from Internal crop research Institute Semi-Arid Tropics,Patnacheru,Hyderabad. seeds surface sterilization 0.01(W/V)inercuric chloride were allowed to germinate at 20oc in dark in petriplates (15 cm in dia) Provided with sterile whatman No.1 filter papers & DW After 72 hr.of incubation five uniform size seedlings were transferred in to each petriplates Provided with whatman No.1 filter papers Each plate contained 10 ml soln of Dw ;500mM NaCl; 500mM NaCl supplimented will

21. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-21 CONT……… This plates were kept in dark roomat20 ± 10c on the 4th day (after transfer into the test solution)5 ml more test soln. was added to the petriplates . Length & fresh wt of the sedlings were recorded on the 7th day . The seedlings were dried in oven at 1100c for 24 hr.& dry wt also recorded. Remark- The BRs not only removed the inhibitory effect of salinity but also promoted the growth in terms of fresh dry wt.

22. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-22 Effect of BRs on salinity induced growth inhibition of groundnut seedlings Compounds Length Fresh weight Dry weight (cm) (gm) (gm) Control (Dw) 7.26 ± 0.32 1.93 ± 0.06 0.40 ± 0.03 500mM Nacl+ 6.35±0.07 2.23 ± 0.07 0.52 ±0.03 1 µM Brassinolide 500mM Nacl+ 6.05±0.17 2.22 ± 0.07 0.48 ±0.01 3µM Brassinolide 500mM Nacl+ 6.28±0.21 2.76 ± 0.09 0.47 ±0.02 1µM 24- epibrassinoloide 500mM Nacl+ 5.94±0.15 2.25 ± 0.09 0.41±0.02 3µM 24- epibrassinoloide 500mM Nacl+ 6.37±0.16 2.42 ± 0.07

23. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-23 Effect of BRs on growth & yield of Tomato The present work is designed to evaluate the influence of exogenously applied brassinosteroides on the growth & economic yield of tomato plants under field condition 24- epibrassinosteroide & 28- homobrassinosteroides were produced from Beak technologies. Inc, Brampton Canada seed of tomato Var. Pusa early dwarf were procured from (NSC) Hyderabad. Seed sown in 4thn week of july 2000 in 100 x 80 cm nursery beds. 20 days old plant were transplanted in expt. Plot 30 x 30cm spacing 1ml of BRs was dissolved in 1ml of ethanol & made up of a vol. 100ml with DW From this stock soln.,0.5,1.0,& 3.0 µM dilution were prepared using DW BRs. Soln. were applied to the plants from plants as a foliar sprayon 35th, 45th,& 55th day after sowing. After 90th day 5 plants were removed for recording the growth in terms of shoot & root length & root & shoot fresh wt. On 120th day fruits were harvested for recording the fruit wt. Yield parameters were considered 1) No. of fruits /plant 2) fresh wt of total fruits /plant.

24. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-24 Treatments control 28- homobrassinolide (3.0µM) 24-Epibrassinolide (3.0µM Shoot length 95.33 ± 3.47 145.66 ± 1.65 144.33 ± 2.12 (cm) Root length (cm) 8.50 ± 0.47 17.60 ±0.52 16.60 ±0.43 No. of fruits /plant 7.00 ± 0.47 14.66 ±0.72 13.66 ±0.72 Total fruit wt /plant 86.66 ± 4.32 220.66 ±6.69 189.0 ±4.58 • Among the three 28- homobrassinoides was most effective in accounting increase the yield. • Present study revealed the ability of BRsin improving the yield of tomato under field condition. • Similar result have also observed in Wheat, (Sairam 1994),Grapes( Xu et al.1994)under field condition. Effect of BRs on growth & yield of tomato

25. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-25 Future prospectus • Blending of two traits –Growth promotion & stress tolerance will have great economic bearing in future agril. Crop production. for • The entry of reputed agrochemical comp.in commercial prodn. of BRs & hence it’s potentiality of improving crop yield can soon be visualized. • The physiological & molecular mechanism of the action of brassinosteroids in enhancing productivity & improving resistance to environmental stress, that’s why it needs further elucidation. • Further research may also bring into focus many more significant roles to the this group of steroids. • New discoveries in this field may allow us to employ BRs as- Highly promising field. Environment friendly natural substance. Wide application in plant protection & yield enhancement. • Availability of BRs at affordable prices will be cost effective. • Expected that chemical promise held by BRs to boost crop production will soon be accomplished.

Brassinosteroids plant harmones

Brassinosteroids plant harmones

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Brassinosteroids plant harmones