Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 103, Issue 2, pp 155–163 | Cite as

Effect of light-emitting diodes on growth and morphogenesis of upland cotton (Gossypium hirsutum L.) plantlets in vitro

  • Huimin Li
  • Zhigang XuEmail author
  • Canming TangEmail author
Original Paper


The objective of this study was to determine the effects of different light-emitting diode (LED) light sources on the growth of upland cotton (Gossypium hirsutum L.) plantlets. Shoot bud apex cuttings of upland cotton (1.0 cm) were transplanted on Murashige and Skoog (MS) basal medium supplemented with 0.1 mg/l 6-benzyladenine (BA) and 0.5 mg/l naphthalene acetic acid (NAA) and cultured in vitro for 45 days. They were exposed to 50 μmol m−2 s−1 photosynthetic photon flux (PPF) and a 12-h photoperiod under six different lights: fluorescent lamp (CON), monochromatic blue LED (B), three blue and red LED mixtures (B:R = 3:1, 1:1, 1:3) and monochromatic red LED (R). The effects of the six light sources on growth and morphogenesis of upland cotton plantlets grown in vitro were investigated. Fresh weight, dry weight, stem length and second internode length were greatest in plantlets cultured under the B:R = 1:1 blue and red LED light, followed by blue LED light, and they were lowest in plantlets cultured under a fluorescent lamp. Chlorophyll content, leaf thickness, palisade tissue length, leaf and stomata area were highest in plantlets cultured under blue LED light. Root activity, sucrose, starch and soluble sugar contents were highest in plantlets cultured under red LED light. Our results showed that larger, healthier plantlets and a greater biomass of upland cotton were produced in the presence of red LED supplemented with a quantity of blue LED light. Blue and red LED (B:R = 1:1) was the most suitable light for the growth of upland cotton plantlets in vitro, and it may be used as alternative light source for an upland cotton culture system.


Blue LED Red LED Upland cotton Plantlet In vitro 



We gratefully acknowledge the technical assistance of Prof Oingya Wang for help in making cross-sections of upland cotton leaves. This work was supported by a grant from 863 National High Technology Program of China (No: 2006AA03A165) and Natural Science Foundation of China (No: 30972035).


  1. Anzelika K, Renata MC, Stase D, Silva Z, Genadij K, Gintautas T, Pavelas D, Arturas Z (2008) In vitro culture of Chrysanthemum plantlets using light-emitting diodes. Cent Eur J Biol 3:161–167CrossRefGoogle Scholar
  2. Arnon DL (1949) Copper enzymer in isolated chloroplast polyphenol oxidase in Beta vulgaris. Plant Physiol 24:1–15CrossRefPubMedGoogle Scholar
  3. Aydin Y, Ipekci Z, Talasora T, Zehir A, Bajrovic K, Gozukirmizi N (2004) High frequency somatic embryogenesis in cotton. Biol Plant 48:491–495CrossRefGoogle Scholar
  4. Bicakci E, Memon AR (2005) An efficient and rapid in vitro regeneration system for metal resistant cotton. Biol Plant 49:415–417CrossRefGoogle Scholar
  5. Bourget CM (2008) An introduction to light-emitting diodes. HortScience 43:1944–1946Google Scholar
  6. Briggs WR, Olney MA (2001) Photoreceptors in plant photomorphogenesis to date, five photochromes, two cryptochrome, one phototropin and one superchrome. Plant Physiol 125:85–88CrossRefPubMedGoogle Scholar
  7. Briggs WR, Beck CF, Cashmore AR, Christie JM, Hunghes J (2001) The phototropin family of photoreceptors. Plant Cell 13:993–997CrossRefPubMedGoogle Scholar
  8. Clouse SD (2001) Integration of light and brassinosteroid signals in etiolated seedling growth. Trends Plant Sci 6:443–445CrossRefPubMedGoogle Scholar
  9. Dewir YH, Chakrabarty D, Hahn EJ, Paek KY (2007) Flowering of Euphorbia millii plantlets in vitro as affected by paclobutrazol, light emitting diodes (LEDs) and sucrose. Acta Hort 764:169–173Google Scholar
  10. Doi M, Shigenaga A, Emi T, Kinoshita T, Shimazaki K (2004) A transgene encoding a blue-light receptor, phot1, restores blue light responses in the Arabidopsis phot1phot2 double mutant. J Exp Bot 55:517–523CrossRefPubMedGoogle Scholar
  11. Duong TN, Hong LTA, Watanabe H, Goi M, Tanaka M (2003) Efficiency of a novel culture system by using light-emitting diode (LED) on in vitro and subsequent growth of micropropagated banana plantlets. Acta Hort 616:121–127Google Scholar
  12. Felker FC, Doehlert DC, Eskins K (1995) Effects of red and blue light on the composition and morphology of maize kernels grown in vitro. Plant Cell Tissue Org Cult 42:147–152CrossRefGoogle Scholar
  13. Hahn EJ, Bae CH, Lee YB (1998) Growth and leaf-surface characteristics of chrysanthemum plantlets between micropropagation and microponic system. J Kor Soc Hort Sci 39:838–842Google Scholar
  14. Hahn EJ, Kozai T, Paek KY (2000) Blue and red light-emitting diodes with or without sucrose and ventilation affects in vitro growth of Rehmannia glutinose plantlets. Plant Biol 43:247–250CrossRefGoogle Scholar
  15. Heo JW, Lee CW, Chakrabarty D, Paek KY (2002) Growth responses of marigold and salvia bedding plants as affected by monochromic or mixture radiation provided by a light emitting diode (LED). Plant Growth Regul 38:225–230CrossRefGoogle Scholar
  16. Hoenecke ME, Bula RJ, Tibbits TW (1992) Importance of blue photon levels for lettuce seedlings grown under red-light-emitting diodes. HortScience 27:427–430PubMedGoogle Scholar
  17. Jao RC, Fang W (2004) Effects of frequency and duty ratio on the growth of potato plantlets in vitro using light-emitting diodes. HortScience 39:375–379Google Scholar
  18. Jao RC, Lai CC, Fang W, Chang SF (2005) Effects of red light on the growth of Zantedeschia plantlets in vitro and tuber formation using light-emitting diodes. HortScience 40:436–438Google Scholar
  19. Jin S, Zhang X, Nie Y, Guo X, Liang S, Zhu H (2006) Identification of a novel elite genotype for in vitro culture and genetic transformation of cotton. Biol Plant 50:519–524CrossRefGoogle Scholar
  20. Kevin W (2000) ‘Photo-Manipulation-Boxes’: an instrument for the study of plant photobiology. Plant Photobiol 26:3–15Google Scholar
  21. Kim SJ, Hahn EJ, Heo JW, Paek KY (2004) Effects of LEDs on net photosynthetic rate, growth and leaf stomata of Chrysanthemum plantlets in vitro. Sci Hort 101:143–151CrossRefGoogle Scholar
  22. Kinoshita T, Doi M, Suetsugu N, Kagawa T, Wada M, Shimazaki K (2001) Phot1 and phot2 mediate blue light regulation of stomatal opening. Nature 414:656–660CrossRefPubMedGoogle Scholar
  23. Kowallik W (1982) Blue light effects on respiration. Plant Physiol 33:51–72CrossRefGoogle Scholar
  24. Kraepiel Y, Mipiniac E (1997) Photomorphogenesis and phytohormones. Plant Cell Environ 20:807–812CrossRefGoogle Scholar
  25. Lee AE, Tewari RK, Hahn EJ, Paek KY (2007) Photon flux density and light quality induce changes in growth, stomatal development, photosynthesis and transpiration of Withania somnifera (L.) Dunal. plantlets. Plant Cell Tissue Organ Cult 90:141–151CrossRefGoogle Scholar
  26. Lian ML, Murthy HN, Paek KY (2002) Effect of light emitting diodes (LEDs) on the in vitro induction and growth of bulblets of Lilium oriental hybrid ’Pesaro’. Sci Hort 94:365–370Google Scholar
  27. Martin AB, Cuadrado Y, Guerra H, Gallego P, Hita O, Martine L, Dorado A, Villalobos N (2000) Differences in the contents of total sugars, reducing sugars, starch and sucrose in embryogenic and nonembrogenic calli from Medicago arborea L. Plant Sci 154:143–151CrossRefPubMedGoogle Scholar
  28. Massa GD, Kim HH, Wheeler RM, Mitchell CA (2008) Plant productivity in response to LED lighting. HortScience 43:1951–1956Google Scholar
  29. Mithilesh K, Bakesh T (2001) Plant regeneration in cotton: a short-term inositol starvation promotes developmental synchrony in somatic embryogenesis. In Vitro Cell Dev Biol Plant 40:294–298Google Scholar
  30. Miyashita Y, Kitaya Y, Kozai T, Kimura T (1995) Effects of red and far-red light on the growth and morphology of potato plantlets in vitro: using light emitting diode as a light source for micropropagation. Acta Hort 393:189–194Google Scholar
  31. Morrow RC (2008) LED lighting in horticulture. HortScience 43:1947–1950Google Scholar
  32. Mortensen LM, Stromme E (1987) Effects of light quality on some greenhouse crops. Sci Hort 33:27–36CrossRefGoogle Scholar
  33. Moshe R, Dalia E (2007) On the effect of light on shoot regeneration in petunia. Plant Cell Tissue Org Cult 89:49–54CrossRefGoogle Scholar
  34. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497CrossRefGoogle Scholar
  35. Nhut DT, Takamura T, Watanabe H, Murakami A, Murakami K, Tanaka M (2002) Sugar-free micropropagation of Eucalyptus citriodora using light-emitting diode (LEDs) and film-rockwool culture system. Environ Control Biol 40:147–155Google Scholar
  36. Nhut DT, Takamura T, Watanabe H, Okamoto K, Tanaka M (2003) Responses of strawberry plantlets cultured in vitro under superbright red and blue light-emitting diodes (LEDs). Plant Cell Tissue Org Cult 73:43–52CrossRefGoogle Scholar
  37. Nhut DT, Takamura T, Watanabe H, Okamoto K, Tanaka M (2005) Artificial light source using light-emitting diodes (LEDs) in the efficient micropropagation of Spathiphyllum plantlets. Acta Hort 692:137–142Google Scholar
  38. Outlaw WH Jr (2003) Integration of cellular and physiological functions of guard cells. CRC Crit Rev Plant Sci 22:503–529CrossRefGoogle Scholar
  39. Ozyigit II, Gozukirmizi N (2009) Efficient shoot and root formation from cotton shoot apices. Ru J Plant Physio 56:527–531CrossRefGoogle Scholar
  40. Puspa RP, Ikuo K, Ryosuke M (2008) Effect of red-and blue-light-emitting diodes on growth and morphogenesis of grapes. Plant Cell Tissue Org Cult 92:147–153CrossRefGoogle Scholar
  41. Saebo A, Krekling T, Appelgren M (1995) Light quality affects photosynthesis and leaf anatomy of birch plantlets in vitro. Plant Cell Tissue Org Cult 41:177–185CrossRefGoogle Scholar
  42. Sakhanokho HF, Peggy OA, May OL, Chee PW (2005) Putrescine enhances somatic embryogenesis and plant regeneration in upland cotton. Plant Cell Tissue Org Cult 81:91–95CrossRefGoogle Scholar
  43. Schuerger AC, Brown CS, Stryjewski EC (1997) Anatomical features of pepper plants (Capsium annuum L.) grown under red light emitting diodes supplemented with blue or far-red light. Ann Bota 79:273–282CrossRefGoogle Scholar
  44. Shimazaki K, Doy M, Assmann SM, Kinoshita T (2007) Light regulation of stomatal movement. Annu Rev Plant Biol 58:219–247CrossRefPubMedGoogle Scholar
  45. Sims DA, Pearcy RW (1992) Response of leaf anatomy and photosynthetic capacity in Alocasia macrorrhiza (Araceae) to a transfer from low to high light. Am J Bot 79:449–455CrossRefGoogle Scholar
  46. Sujatha M, Sailaja M (2005) Stable genetic transformation of castor (Ricinus communis L.) via Agrobacterium tumefaciens-mediated gene transfer using embryo axes from mature seeds. Plant Cell Rep 23:803–810CrossRefPubMedGoogle Scholar
  47. Sulekha H, Anuradhav K, Satishm N, Anjank B, Dineshc A, Kazav K (2000) Influence of explants, genotypes and culture vessels on sprouting and proliferation of pre-existing meristems of cotton (Gossypium hirsutum L. and Gossypium arboreum L.). In vitro Cell Dev Biol Plant 36:505–510CrossRefGoogle Scholar
  48. Sun YQ, Zhang XL, Huang C, Guo XP, Nie YC (2006) Somatic embryogenesis and plant regeneration from different wild diploid cotton (Gossypium) species. Plant Cell Rep 25:289–296CrossRefPubMedGoogle Scholar
  49. Sun JY, Li WM, Zhang HS, Zhao JL, Yin XL, Wang L (2009) Somatic embryogenesis and plant regeneration in glandless upland cotton (Gossypium hirsutum L.). Front Agric China 3:279–283CrossRefGoogle Scholar
  50. Takahashi K, Fujino K, Kikuta Y, Koda Y (1995) Involvement of the accumulation of sucrose and the synthesis of cell wall polysaccharides in the expansion of potato cells in response to jasmonic acid. Plant Sci 111:11–18CrossRefGoogle Scholar
  51. Tanaka M, Takamura T, Watanabe H, Endo M, Yanagi T, Okamoto K (1998) In vitro growth of Cymbidium plantlets cultured under super bright and blue light-emitting diodes (LEDs). J Hort Sci Biotech 73:39–44Google Scholar
  52. Tanreer K, Akii KS, Pant RC (2006) Regeneration via somatic embryogenesis and organogenesis in different cultivars of cotton (Gossypium spp.). In Vitro Cell Dev Biol Plant 42:498–501CrossRefGoogle Scholar
  53. Tripathy BC, Brown CS (1995) Root-shoot interaction in the greening of wheat seedlings grown under red light. Plant Physiol 107:407–411PubMedGoogle Scholar
  54. Wang QY (1982) Microscope technology of plant. Plant Teaching and Research Group of Nanjing Agricultural University, Nanjing (in Chinese)Google Scholar
  55. Wongnok A, Piluek C, Techasilpitak T, Tantivivat S (2008) Effects of light emitting diodes on micropropagation of Phalaenopsis orchids. Acta Hort 788:149–156Google Scholar
  56. Zeng B, Wang QY, Tang CM (2008) Anatomic analysis on heterosis in three transgenic bt pest-resistant hybrid cotton (G. hirsutum L.). Acta Agron Sin 34:496–505 (in Chinese)CrossRefGoogle Scholar
  57. Zhang BH, Liu F, Liu ZH, Wang HM, Yao CB (2001) Effects of kanamycin on tissue culture and somatic embryogenesis in cotton. Plant Growth Reg 33:137–149CrossRefGoogle Scholar
  58. Zhang YS, Huang X, Chen YF (2009) Experimental course of plant physiology. Higher Education Press, Beijing (in Chinese)Google Scholar
  59. Zhao HM, Ai HL, Wei CY (2007) Effects of hygromyc in on cotton cultures and its application in agrobacterium-mediated cotton transformation. In Vitro Cell Dev Biol Plant 43:111–118CrossRefGoogle Scholar
  60. Zhu SW, Sun JS (2000) Rapid plant regeneration from cotton (Gossypium hirsutum L.). Chin Sci Bull 45:1171–1174Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.College of Agronomy, National Key Laboratory of Crop Genetics & Germplasm EnhancementNanjing Agricultural UniversityNanjingPeople’s Republic of China

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