Ignacio Porras

Physiological and growth changes in micropropagated Citrus macrophylla explants due to salinity.

Salinity is one of the major abiotic stresses affecting arable crops worldwide, and is the most stringent factor limiting plant distribution and productivity. In the present study, the possible use of in vitro culture to evaluate the growth and physiological responses to salt-induced stress in cultivated explants of Citrus macrophylla was analyzed. For this purpose, micropropagated adult explants were grown in proliferation and rooting media supplemented with different concentrations of NaCl. All growth parameters were decreased significantly by these NaCl treatments; this was accompanied by visible symptoms of salt injury in the proliferated shoots from 60mM NaCl and in the rooted shoots from 40mM NaCl. Malondialdehyde (MDA) increased with increasing salinity in proliferated shoots, indicating a rising degree of membrane damage. The concentration of total chlorophyll significantly decreased in the presence of NaCl, and this effect was more pronounced in the rooted explants. The Na(+) and Cl(-) concentrations in the explants increased significantly with the salinity level, but Cl(-) levels were higher in the proliferated explants than in the rooted explants. For osmotic adjustment, high concentrations of compatible solutes (proline and quaternary ammonium compounds-QAC) accumulated in salt-stressed plants in proliferation, but differences were not observed in rooted explants. In proliferation, proline and QAC were highly correlated with the sodium and chloride concentrations in the explants, indicating a possible role of these compounds in osmotic adjustment. The plant concentrations of NO(3)(-), K(+), Mg(2+), Ca(+) and Fe were also affected by the NaCl concentration of the medium. We suggest that the important deleterious effects in the in vitro explants of Citrus macrophylla grown at increasing NaCl concentrations were due mainly to toxic effects of saline ions, particularly Cl(-), at the cellular level.

Fruit splitting in 'Nova' hybrid mandarin in relation to the anatomy of the fruit and fruit set treatments

Fruit splitting in the mandarin orange cultivar Nova is related to the presence of an open stylar end in the fruit. This opening arises from a cavity present at the bas e of the style, and is formed in a variable proportion of fruits. The cells at the bottom of this style cavity form a rind-like tissue which develop into a navel structure. The size of this navel is larger in open-stylar end bearing fruits and, accordingly, in split fruit. The number of split fruit per tree is unrelated to the number of fruit set. The application of gibberellic acid ( GA3 ) has a variable effect on set and splitting depending on its timing. This hormone increases fruit splitting when applied at flowering, but reduces it when applied shortly after the end of the June drop. It has only a minor effect on fruit set.

Efficient propagation and rooting of three citrus rootstocks using different plant growth regulators

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 N6-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.

High efficiency in vitro organogenesis from mature tissue explants of Citrus macrophylla and C. aurantium

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 N6-benzyladenine (BA) concentrations or with 2xa0mgu2009l−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 2xa0mgu2009l−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 4xa0wk was essential for regeneration process in both rootstocks.

Quality and fruit colour change in Verna lemon

While most of lemon cultivars in the northern hemisphere are harvested in autumn-winter, Verna, an autochthonous Spanish cultivar, is harvested later (February to August), supplying the European market when lemons are in short supply, a market that is also served by imports from the southern hemisphere, mainly Argentina and South Africa. The aim of this study was to determine the temperature at which degreening begins naturally in Verna lemon, noting the evolution of colorimetric parameter a and comparing the same with the equivalent measurements made in Eureka lemon, the most widely cultivated lemon worldwide. The influence of solar radiation on the colorimetric parameters was studied, and then the influence of the minimum temperatures on the change from green to yellow, using the data collected over five growing seasons, was also assessed. The results confirmed the relation between net solar radiation and degreening in Verna, a process that begins when net solar radiation reaches a value of between 2 and 4 MJ/m 2 .day and when the mean temperature of the 14 days prior to sampling is8.8 oC or when the daily mean temperature reaches5.5 oC on two consecutive days. The information obtained will enable growers to predict the colour changes that will occur in the field and potential growers to ascertain whether a given geographical zone is suitable for the crop in question.