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Changes in morphology and biochemical indices in browning callus derived from Jatropha curcas hypocotyls

Abstract

Callus browning is a typical feature of callus cultures derived from the hypocotyl of Jatropha curcas. Brown callus results in decreased regenerative ability, poor growth and even death. In this study, we investigated the effect of browning on callus morphology and biochemical indices. Light microscopy and scanning electron microscopy showed striking differences in callus morphology. During browning, chlorophylls and carotenoids concentrations decreased steadily. Polyphenol oxidase (PPO) and peroxidase (POD) enzymatic activities patterns were similar during callus culture with a higher activity level at week 3 compared to week 2 or later weeks. Grey relation degree analysis indicated that PPO played a more important role than POD in enzymatic callus browning. Polyacrylamide gel electrophoresis results showed differences between browning and non-browning callus. Gas chromatography–mass spectrometry results showed that saturated and unsaturated fatty acid quantities differed significantly but there was little difference in fatty acid composition between non-browning and browning callus. Differences in 17, 18.4 and 25 kDa protein concentrations were also observed in browning and non-browning callus using sodium dodecyl sulfate–polyacrylamide gel electrophoresis.

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Abbreviations

LM:

Light microscopy

SEM:

Scanning electron microscopy

BA:

Benzylaminopurine

IBA:

Indole-3-butyric acid

MS:

Murashige and skoog medium

PPO:

Polyphenol oxidase

POD:

Peroxidase

PBS:

Phosphate buffer solution

SDS:

Sodium dodecyl sulfate

PAGE:

Polyacrylamide gel electrophoresis

d:

Day

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Acknowledgments

We would like to thank Yongjiong Jia, Shenghua Wang, and Lin Tang for their invaluable suggestions. This work was supported by the grants from [“Eleventh Five Years” Key Project of Chinese Minisity of Science and Technology (no. 2006BAD07A04 and no. 2007BAD50B05); and Key Project of Chinese Minisity of Education (no. 307023 and no. 20060610015)].

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Correspondence to Fang Chen.

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He, Y., Guo, X., Lu, R. et al. Changes in morphology and biochemical indices in browning callus derived from Jatropha curcas hypocotyls. Plant Cell Tiss Organ Cult 98, 11–17 (2009). https://doi.org/10.1007/s11240-009-9533-y

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Keywords

  • Jatropha curcas
  • Browning
  • Callus
  • PPO
  • POD
  • Tissue culture