Bioluminescence in insects

bioluminous insects and its features

1. welcome

2. BIOLUMINESCENCE IN INSECTS Gande sravan kumar Roll No:M-1063/17

3. Introduction History of Bioluminescence Significance of Bioluminescence Survey of Luminous insects Physiology of light emission in Lampyridae Chemistry of Bioluminescence in Lampyridae Applications of Bioluminescence Current projects Case studies Conclusion

4. Introduction

5. Cont… Cold light emission Hybrid word, Greek → Bios = Living Latin → Lumen = Light Also known as ‘ Chemiluminescence ’ .

6. Seen in Jelly fish Marine octopus Panellus stipticus )

7. Unique – color, wavelength, timing & interval Throughout the biosphere, but only at levels below the mammals & plants 17 phyla & 700 genera Terrestrial organisms (Annelids, Arthropods, Molluscs ), Fungi, and microorganisms

8. Insects have received much attention

9. History of Bioluminescence

10. Greeks and Romans - characteristics of luminous organisms 1555 – first book devoted to Bioluminecsence – Conrad Gesner 1667 – Robert Boyle – air requirement 1873 - Raphael Dubois – extracted & coined the terms " luciferine " and "luciferase". 1959 – Harvey – Luciferin – luciferase system Cont…

11. Significance ( Gajendra Babu & Kannan ., 2002 )

12. Eg :- Fireflies Same species to aggregate Two communication systems : 1. wingless & sedentary females( Lampyris spp ) 2. Photinus spp & Photuris spp Male fireflies seeking mates Female : male – 1 : 50 Flash pattern vary with species & sex Flashes begin / end sharply Mating signal

13. Photuris pyralis :- ( Llod ., 1971 , Sivinsky , 1981)

14. New Zealand glow worm fly ( Arachnocampa luminosa ( Diptera : Keroplatidae ) Female fly – egg – ceiling of dark caves in Waitomo Prey Eg :- Chironomid midges Luminous caves ( tourist attraction spots in New Zealand ) Show route ( Gajendra Babu & Kannan ., 2002 ) Predation

15. Photuris spp : Female imitate pattern of females of other species (mostly P hotinus spp.) Lure males of those species Feed on them ( femme fatale) ( Vencl et al., 1994 )

16. Eg :- Rail road worms (Order: Coleoptera ) Lateral light organs Live in high densities Sudden flashes Repel potential predators Intimate mated females – lay eggs – over crowding & competition for food source ( Gajendra Babu & Kannan ., 2002 ) Defence

17. Eg :- Rail road worms Head light organs Red light ( long wavelength ) Not visible to prey Search for millipedes ( Viviani & Bechara ., 1997 ) Illumination

18. Survey COLLEMBOLA (spring tails) DICTYOPTERA ( cockroaches ) HEMIPTERA Fulgoridae – lantern flies DIPTERA ( Glow worm flies ) Bolitophilidae Platyuridae COLEOPTERA ( Beetles ) Elateridae - click beetles Lampyridae - fireflies Phengodidae - railroadworms

19. c Adult Eg :- Onychiurus armatus Soil Greenish flash throughout the body Fat body Stimulation is required Significance unknown (rare) Collembola Eg : Giant cockroach ( Blaberidae ) Luchhormetica luckae Recently found in the Amazon forest. It mimics click beetle ( Peter Vršanský et al 2012)   DICTYOPTERA Fulgora laternaria (F :- Fulgoridae ) Adults Both sexes should be together Mating signal No work on physiology & biochemistry Hemiptera : lantern flies

20. Diptera Glow worm flies - F:- Bolitophilidae Eg :- Arachnocampa luminosa or Bolitophila luminosa Excretory organ – photogenic function Larvae – blue to green Female – weakly luminescent Male – loose after emergence from pupae Eg :- Orfelia fultoni F : Mycetophilidae Larva – Blue color Webs – along stream banks in the Appalachian mountains Lure prey Light organ – anterior part & in tail

21. Eg :- K eroplatus sesoides Real fungus gnat Eat fungal spores (Oospores) No special light organ Hypodermal fat body – stimulated Whole body of larva & pupa is luminou s

22. COLEOPTERA ( Beetles ) : Most of the bioluminescent species Super family :- Elateroidea F :- Lampyridae ( fireflies & glow worms ) Larvae & adult – luminous Yellow – green flashes – ventral lanterns Species – specific signal ( adult ) Aposematism ( warning signal ) – larvae – abdominal lanterns

23. F :- Elateridae ( click beetles ) Green glow – prothoracic lanterns – disturbed – defense Yellow – orange glow – abdominal lanterns – flying Luminous termite mounds – Brazilian savanna spp (attract flying prey )

24. F :- Phengodidae ( R ailroadworms ) Male – not luminous Larvae & female – exuberant Phrixothrix spp Lateral lanterns – yellow green Head lanterns – red Only terrestrial organism producing red light

25. Variation in Colour of Light Varies with species, environmental factors or structure of luciferase Collembola – greenish flash throughout the body Fulgora ( Hemiptera ) _ white Most insects - yellow-green as in Photinus and Lampyris ( Coleoptera ). In larval and adult female railroad worms, thorax and abdomen – green to orange. head - red light Arachnocampa - blue-green

26. Physiology of light emission in Lampyridae Light organ – photophore / lantern May occur in both sexes Eg :- Fire flies Photuris spp ( ventral side ) Males -2pairs Female -1pair

27. Rail road worms : (larvae & female ) 11 pairs- dorso lateral side of thorax and abdomen 1 pair on head Fulgora - 1 light organ on head

28. Simplest case :- Eg :- Phengodid larvae Gaint oenocyte like cells Lack neural control ( slow responding light system ) Specialized case :- Eg :- Photinus spp & Photuris spp Rousettes of thousands of photocytes interpenetrated by tracheaeoles

29. Photocytes – lie beneath epidermis Cuticle – transparent ( permit light ) Trachea and nerves perpendicular to cuticle Tracheal end cell – sphincter ( neuralcontrol ) Tracheolar cell Neuron – spatulate terminal processes

30. 32

31. 3 substances are involved (Raphael Dubois., 1885 ) Luciferin – heat resistant substrate & source of light Luciferase – enzyme – trigger Molecular oxygen – fuel ATP, Mg+2 ions – catalysts Chemical reaction occurs in cytoplasm Chemistry

32. Luciferin – low molecular weight compound – an aldehyde, a polypeptide complex or a protein Sequence of events involved in light production in insects

33. Importance Besides their beauty and scientific interest - very important to the society luciferases and luciferins - sensitive tools - biotechnology and biomedicine. luciferase genes - bioluminescent markers - tuberculosis and HIV Biosensors - pollutants and environmental disruptors

34. Applications Space Research Pest management Fluorescent marker gene Medical research

35. Space Research Luciferin & luciferase systems – spacecrafts – Mars Existence of life forms Special electronic device Pick up soil Mix with water, oxygen, luciferin & luciferase Glow ATP ( 5 th requirement ) one quadrilionth of a gm - sensitive

36. Pest management Organisms distribution pattern USA, 2001 – GFP – jelly fish ( Aequora victoria ) Modified genetic material of Pink boll worm Transgenic PBW strain ( green flourescent larvae ) Objective – 2 fold 1. Distribution pest 2. Temperature sensitive lethal gene ( management )

37. Fluorescent marker gene Marker gene – short sequence of DNA Acts as a label Inserted along with a gene of interest into cells Fluorescent marker genes – transformed cells glow under light Help scientists – select transformed cells

38. Medical research Luciferase is injected into cells which glow Monitor progress in treatment as cells glowing begin to disappear Muscular dystrophy, heart disease, urology, antibiotic testing and waste water treatment Blood banks have used the luciferase enzyme to test the quality of red blood cells within their stock.

39. How Luciferin Help Us Understand Cancer Normal cells Respond to growth factors ( signals ) Food & nutrients – instructions Excess growth & cell division is avoided Cancer cells Ignore signals Uncontrolled uptake of food & nutrients due to mutation in genes Tumor formation

40. 1924, Otto Warburg Glucose – cancer cells > normal cells Convert to Lactate Plenty of oxygen – aerobic glycolysis Altered metabolism – Warburg effect Tumor cells produce lactate even when oxygen is present

41. The firefly "Christmas Tree" as observed in regions of S.E Asia, from Malaysia to Papua-New Guinea. It is due to the synchronous flashing of fireflies.

42. Image of bioluminescent red tide event of 2005 at a beach in Carlsbad California showing brilliantly glowing crashing waves containing billions of Lingulodinium polyedrum dinoflagellates . The blue light is a result of a luciferase enzyme .

43. Current projects Structure / Function relationship of insect luciferase Molecular origin and evolution of insect luciferase Biodiversity and photoecology of terrestrial bioluminescence Prof. Dr. Vadim Viviani, Departamento de Biologia, Brazil.

44. Construction, characterization and exemplificative application of bioluminescent Bifidobacterium longum ( Guglielmetti s. et al, 2008). 1 location: Department of Food Science and Microbiology, University of Milan, Italy Institute of Environmental Engineering and Biotechnology,Tampere University of Technology, Finland

45. AIM : To construct a bifidobacterial luminescent biosensor that could be used for a quick analysis of the metabolic state of cells under different conditions. RESULTS : Detected a minimum of 4000 cells, which indicates that the insect luciferase expression in Bifidobacterium longum is extremely good, and a measurement requires only a few minutes of incubation

46. 2 ATP-Bioluminescence as a method to evaluated microbiological quality of UHT milk ( A.F. Cunha et al,2014). LOCATION : Brazil United States – St. Paul, MN, United States.

47. AIM : To compare the results of culturemethods with the results of ATP-Bioluminescence technique of 102 UHT whole milk samples incubated at 48, 72, and 168 hours. RESULT : Analyzed for the presence of mesophilic and psychrotrophic aerobic microorganisms using Plate Count Agar (PCA), Brain-Heart Infusion (BHI)media and PetrifilmTM Aerobic Count (AC) plate. For the dairy industry, the ATP-Bioluminescence techniquemay become an important tool that assists the official methods to quickly monitor the microbiologicalquality of UHT milk though this will likely require a threshold below 150 RLU.

48. Conclusion :- Bioluminescence - light by a living organism ( cold light ) 2000 lumiscent species in insects Functional significance ( mating signal, defence , predation ) – biological control of pests Luciferase – sensitive tool in biotechnology, biomedicine Biosensors – pollutants & environmental disruptors