The quality of acclimatized in vitro cultivated plants is essential to ensure good survival and growth after planting in field conditions. After two months of acclimatization, this study revealed a significant variation in survival rate, shoot characteristics, root traits, and biomass features between 13 clones of Melia volkensii Gürke. A number of quality indices such as the Dickson Quality Index (DQI), shoot dry weight: root dry weight ratio (S:R), and sturdiness quotient (SQ) also showed a large variation. The survival rate was genotype-dependent, with an average of 85%. Extreme genotypes were tall, had long internodes, a lot of leaves, and a large leaf area. At the other side of the spectrum, there were small clones with short internodes and a reduced number of leaves and leaf areas. The high S:R values (>2) of the evaluated clones indicate an imbalance between the shoot and root system, negatively influencing the quality of the in vitro produced M. volkensii plants. The SQ for all clones was below the threshold value of 6, implying a good and expected survival rate. This study illustrates that some clones might systematically score better for SQ, S:R, plant volume, and DQI after acclimatization and that these factors could form the basis for selection.

Before in vitro propagated Melia volkensii plants can be used for mass planting, the transition phase to in vivo conditions needs to be better controlled because too many plants are lost during acclimatization and in the field. Two experiments were set up to evaluate the effects of biological agents on the establishment of M. volkensii in vitro plantlets. The biological agents consisted of Trichotech®, Bio-cure B®, Rhizatech®, Bacillus subtilis, a Trichoderma isolate and self-isolated native arbuscular mycorrhizal fungi (AMF). Regarding the latter, in soil from the nursery, the number of AMF spores increased from six spores to 400 per 100 g of soil using a trap culture, in which thirteen AMF morphotypes were identified and root colonization assessed through observation of hyphae, vesicles, coils and appressoria. The first experiment was set up in the greenhouse to investigate the efficacy of the biological agents on the hardening off. In the second, a field experiment was set up to study their effect on the early establishment of the plantlets in the field compared to seedlings. All biological agents significantly (p ≤ 0.05) improved in vitro plant survival and growth compared to the control. The highest plant height and number of leaves per plant were recorded in plants treated with Rhizatech®, Native AMF, Bio-cure B® and Trichoderma isolate. The treatments with Rhizatech®, Bio-cure B® and native mycorrhiza recorded a significantly wider stem. The root diameter of the plants treated with Rhizatech® and Bio-cure B® was the largest, but the plants inoculated with the native AMF had the longest roots. Moreover, the inoculated plants generally developed multiple secondary roots. After two months, AMF had clearly colonized the acclimatized plantlets. In the field experiment, the biologicals made no difference in survival rate but did produce a significantly larger leaf area after two months, with the largest leaves recorded with Rhizatech®, native AMF and Trichotech®. They also increased the quality index of the plants from 0.21 to 0.52. The performance of in vitro grown M. volkensii plants six months after planting in semi-arid conditions in Kiambere was better than that of seedlings. Inoculation of plants increased plant height and diameter. Thus, inoculation of biological agents is an efficient approach for improving the early growth of in vitro propagated M. volkensii plants.

Polyploidy was induced in Melia volkensii (Mukau), a valuable native tree from the semi-arid regions of East Africa. Cotyledons of diploid M. volkensii (2n = 2x = 28) were treated with oryzalin for 0 (control), 1, 2, or 3 h with or without pretreatment with 1.1 µM thidiazuron. Cotyledons treated with 10 mg·L−1 oryzalin for three hours yielded 40% tetraploids. Pretreatment of cotyledons with thidiazuron for 18 days followed by treatment with oryzalin increased tetraploid plant production to 52.5%, but this also yielded more mixoploids. Compared to diploid M. volkensii, the tetraploid in vitro and young potted plants were compacter, with thicker stems, wider leaves, and a low density of longer and wider stomata. In the coming years, tetraploid M. volkensii plants will be observed in field trials and serve as a basis for further breeding efforts.

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