Matching-typing complementarity in histocompatibility testing.

As part of the effort to develop optimal plant varieties for the production and molecular farming of plant-made pharmaceuticals, this study evaluated shoot organogenic potential of a total of 115 Nicotiana accessions, representing 53 species. To induce shoots, leaves from seedling grown in vitro were cut into pieces, cultured on shoot-induction medium under low light for 3 weeks, and then subcultured onto the same medium for another 4 weeks under normal light. Statistical analysis detected significant differences among the 115 accessions for the percentage of leaf explants producing shoots and the number of shoots produced per responsive leaf explant. Importantly, regression analysis also found an exponential relationship between the number of shoots produced per responsive leaf explant and the percentage of leaf explants producing shoots. The number of shoots produced per responsive leaf explant increased rather slowly, ranging from zero to around five, as the percentage of leaf explants producing shoots increased from 0 to 80%, but the increase became dramatic as the percentage increased from 80% to 100%, reaching as high as 35 shoots per responsive leaf explant. This exponential relationship is the first of its kind to be established in plant regeneration studies using either organogenesis or somatic embryogenesis systems. A possible mechanism that governs the establishment of the exponential relationship is discussed.

Culture Techniques↗

PubMed2003

Roles for auxin during morphogenesis of the compound leaves of pea ( Pisum sativum).

The goal of this study was to explore the impact of the plant growth regulator auxin on the development of compound leaves in pea. Wildtype ( WT) plantlets, as well as those of two leaf mutants, acacia ( tl) and tendrilled acacia ( uni-tac) of pea ( Pisum sativum L.), were grown on media containing the auxin-transport inhibitors 2,3,5-triiodobenzoic acid (TIBA), N-(1-naphthyl)phthalamic acid (NPA), or the auxin antagonist, p-chlorophenoxyisobutyric acid (PCIB). The resulting plantlets were carefully analyzed morphologically, by scanning electron microscopy and for Uni gene expression using quantitative reverse transcription-polymerase chain reaction. Auxin transport was measured in WT leaf parts using [(14)C]indole-3-acetic acid. Relative Uni gene expression was determined in shoot tips of a range of leaf-form mutants. Morphological abnormalities were observed for all genotypes examined. The terminal tendrils on WT plants were converted to leaflets, stubs or were aborted. The number of pinna pairs produced on leaves was reduced, with the distal forms being eliminated before the proximal ones. Some leaves were converted to simple, including tri-and bilobed, forms. These treatments phenocopy the uni-tac and unifoliata ( uni) mutants of pea. In the most extreme situations, leaf blades were completely lost leaving only a pair of stipules or scale leaves. Polar auxin transport was basipetal for all leaf parts. Uni gene expression in shoot tips was significantly reduced in 60 microM NPA and TIBA. Uni mRNA was more abundant in tl, af and af tl and reduced in the uni mutants compared to WT. These results indicate that an auxin gradient plays fundamental roles in controlling morphogenesis in the compound leaves of pea and specifically it: (i). is the driving force for leaf growth and pinna determination; (ii). is necessary for pinna initiation; and (iii). controls subsequent pinna development.

Culture Techniques↗

PubMed2003

Regeneration of flax ( Linum usitatissimum L.) plants from anther culture and somatic tissue with increased resistance to Fusarium oxysporum.

The aim of this study was to establish a protocol for the efficient production of flax plants of microspore origin. The results were compared to those obtained for plants regenerated from somatic explants from hypocotyls, cotyledons, leaves, stems and roots. All the plants obtained during the experiments were regenerated from callus that was grown for periods from a few weeks to a few months before the regeneration was achieved. Anther cultures were less effective in plant regeneration than somatic cell cultures. However, regenerants derived from anther cells showed valuable breeding features, including increased resistance to fungal wilt. The age of the donor plants and the season they grew in had a noticeable effect on their anther callusing and subsequent plant regeneration. Low temperature had a negative effect and dark pre-treatment a positive effect on callusing and plant regeneration. Different media were most effective for callus induction, shoot induction and rooting. For callus induction two carbon sources (2.5% sucrose and 2.5% glucose) were most effective; for shoots, only sucrose at lower concentration (2%) was effective. Rooting was most efficient in 1% sucrose and reduced (50%) mineral concentration in the medium. It was found that the length of in vitro cultivation significantly increases the ploidy and affects such features as regenerant morphological characteristics, petal colour, and resistance to Fusarium oxysporum-induced fungal wilt. The established plant regeneration system provides a basis for the creation of transgenic flax.

Culture Techniques↗