In Vitro Propagation of Citrus Rootstocks

Notulae Botanicae Horti Agrobotanici Cluj-Napoca 01/2009;
Source: DOAJ

ABSTRACT Present investigation was conducted to standardize a protocol for in-vitro propagation of citrus rootstocks viz. Rough lemon, Cleopatra mandarin Pectinifera and Troyer citrange. The shoot tip explant was found better for callus induction of these rootstocks than the nodal segment. Maximum callus formation (40.0% and 23.3%) of shoot tip explants was obtained in Cleopatra mandarin, Pectinifera, and Rough lemon and Troyer citrange, respectively in treatment MS basal media + 0.5mg/l Kin, 2.0mg/l NAA, and 2.0mg/l 2, 4-D. Furthermore, the maximum number of shoots per explant was obtained through the callus in Pectinifera, Rough lemon and Cleopatra mandarin in MS basal media + BAP 1mg/l. Maximum rooting of shoots (1.11%) was noted in rootstock Rough lemon followed by Cleopatra mandarin for the ½ MS media supplemented with 10mg/l IBA. Although the callus development and bud proliferation was recorded in rootstock Troyer citrange however, shoot and root formation did not occur. The potting media consisting of soil, sand and FYM in the ratio of 1:1:1 by volume was better with maximum survival rate of hardened plants six weeks after transferring to the pots under greenhouse for Rough lemon followed by Pectinifera and Cleopatra mandarin rootstock.

  • [Show abstract] [Hide abstract]
    ABSTRACT: A simple and efficient protocol for obtaining organogenesis from mature nodal explants of Citrus macrophylla (alemow) and Citrus aurantium (sour orange) has been developed by optimizing the concentrations of the plant growth regulators, the incubation conditions, the basal medium and by the choice of explant. In order to optimize the plant growth regulator balance, explants were cultured in the regeneration medium supplemented with several N 6-benzyladenine (BA) concentrations or with 2 mg l−1 BA in combination with kinetin (KIN) or 1-naphthaleneacetic acid (NAA). The presence of BA was found to be essential for the development of adventitious buds; the best results were obtained using BA at 3 and 2 mg l−1 for alemow and sour orange, respectively. The combination of BA with KIN or NAA in the culture medium decreased the regeneration frequency, with respect to the use of BA alone. The effect of three different basal media was rootstock-dependent. For C. macrophylla the best results were obtained with Woody Plant Medium or Driver and Kuniyuki Walnut Medium (DKW). However, for C. aurantium, although high percentages of regenerating explants were obtained independently of the basal medium used, the highest number of buds per regenerating explants was obtained with DKW medium. Attempts were made to identify the type of explants which had a higher regeneration ability using particular regions along the mature shoots of C. macrophylla. When nodal segments, where the buds were completely removed, and internode segments were compared, the highest percentage of responsive explants was obtained with nodal segments. The existence of a morphogenetic gradient along the shoot was observed and the organogenic efficiency was highest when explants from the apical zone were used. Incubation in darkness for 3 or 4 wk was essential for regeneration process in both rootstocks.
    In Vitro Cellular & Developmental Biology - Plant 04/2012; 49(2). DOI:10.1007/s11627-012-9476-6 · 1.16 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Citrus jambhiri (rough lemon) is considered a major rootstock source for a number of Citrus species. A simple method for micropropagation from nodal segments is reported. Nodal segments of C. jambhiri were inoculated on MS medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP), kinetin, and N6-(2-isopentenyl) adenosine (2iP). Maximum multiple shoot regeneration response (75 %) was observed with BAP at 3 mg l−1. Shoots were multiplied for 30 d on fresh medium with similar composition. A total of 67 % of the cultures showed multiplication with the optimum number of shoots (4.02) and height of shoots (1.81 cm) with BAP (3 mg l−1) alone. Maximum rooting response (87 %) was observed with naphthaleneacetic acid at 0.5 mg l−1. Transverse sections of shoot stems obtained in vivo (sampled from seedlings) and in vitro (regenerated from nodal segments), showed similar anatomies. Randomly amplified polymorphic DNA analysis confirmed that all the regenerated plants were genetically identical to their donor plant, suggesting absence of detectable genetic variation in the regenerated plantlets.
    In Vitro Cellular & Developmental Biology - Plant 10/2012; 48(5). DOI:10.1007/s11627-012-9430-7 · 1.16 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The influence of various basal medium and plant growth regulators on the efficient micropropagation of nodal explants from mature trees of alemow, sour orange, and ‘Cleopatra’ mandarin citrus rootstocks was studied. All three citrus rootstock shoot cultures showed a preference for high-salt media, like Murashige and Skoog or Driver and Kuniyuki Walnut medium. Several combinations of N 6-benzyladenine (BA) and adenine (AD), kinetin (KIN) or gibberellic acid (GA) were tested to optimize the shoot proliferation phase. BA/GA combinations improved the proliferation of all the rootstocks studied, especially alemow. The addition of BA and AD to the culture medium improved shoot proliferation in sour orange and ‘Cleopatra’ mandarin in the same way as BA and GA. The addition of different combinations of BA/KIN did not result in further improvement of any of the studied variables. The transfer of in vitro shoots to rooting media, containing different concentrations of indolebutyric acid (IBA) and indoleacetic acid (IAA), resulted in regeneration of complete plantlets. Alemow and ‘Cleopatra’ mandarin shoots rooted well using these plant growth regulators; however, all combinations of IBA and IAA tested resulted in very low rooting percentages in sour orange. To improve rooting in sour orange and ‘Cleopatra’ mandarin, different combinations of naphthaleneacetic acid (NAA) and IBA were tested. All NAA/IBA combinations produced higher rooting percentages than did the IBA/IAA combinations, and in sour orange nearly 100 % of explants developed roots. An efficient and simple protocol for the micropropagation of three citrus rootstocks, alemow, ‘Cleopatra’ mandarin, and sour orange, by culturing nodes from mature plants, has been established.
    In Vitro Cellular & Developmental Biology - Plant 10/2012; 48(5). DOI:10.1007/s11627-012-9457-9 · 1.16 Impact Factor


Available from