Validating Field Regeneration Capacity for Selected Accessions of Gossypium hirsutum Using Callus Induction and Regeneration Capacity

Gossypium hirsutm undergoes a rapid clonal propagation to regenerate a mature plant through tissue culture. In this research, cotton leaf regeneration level for 21 accessions in the field (new leaves) was observed after the first harvest, and a comparison between field regeneration level and callus induction with its regeneration capacity (new shoots and roots) for the same 21 accessions was carried out. During the flowering stage of Gossypium hirsutum, biochemical (Proline), physiological (chlorophyll and carotenoid content) analysis was carried out. Phenotypic observations (plant height, leaf area, fresh leaf weight, dry leaf weight, flower and boll number) were also carried out on 21 accessions for each accession. Callus induction and regeneration capacity of roots and shoots for hypocotyl, cotyledons and shoot tip tissues was used to validate field regeneration capacity through analysis of variance. ZS061, LuMian378, JiMian863, and ZS065 have highest drought tolerance while ZhongMianSuo24, LiaoYangDuoMaoMian, and BeiZheGongSheMian have the lowest tolerance to drought stress. Accessions with both field and callus regeneration capacity were identified.

Predominance of clonal propagation conceals extinction risks of the highly endangered floodplain herb Cnidium dubium

Habitat loss and degradation due to human-induced landscape alterations are considered to be a major threat to biodiversity. The decline of biodiversity may occur with a time delay leading to a so called extinction debt. Therefore, determining extinction risks and conservation status is not always straightforward. Several life history traits might play a role for the accumulation of an extinction debt. Thus, perennial plant species capable of vegetative propagation might be able to persist temporarily in degraded habitats even though sexual and evolutionary processes are effectively halted. We studied Cnidium dubium, which occurs in scattered patches along river corridors in Central Europe and is critically endangered in Germany. It is a perennial species which is able to propagate clonally. Our aims were to reconstruct demographic processes regarding clonal propagation and gene flow along 400 km of river stretch and with respect to the position in the flooplain, i.e. before or behind dykes. We also wanted to determine whether there is evidence for an extinction debt in C. dubium and to use our insights for conservation recommendations. For this, we used nuclear microsatellites and maternally inherited chloroplast DNA markers and applied a systematic grid based sampling strategy for small scale geographic structures. We observed a high level of clonal propagation. In 935 analysed plants we observed only 121 different genotypes and of 50 studied patches of C. dubium the majority (31 patches) consisted of one single genotype each. Patch size and position were correlated with clonal diversity. Large patches and patches behind dykes exhibited higher clonal diversity. There was no evidence for a large scale genetic substructuring of the study area and no differences in overall genetic diversity between upstream and downstream patches as well as between patches before and behind the dykes. High levels of heterozygosity and a high number of 18 chloroplast DNA haplotypes togetherwith a slightly elevated inbreeding coefficient (Fis) point toward a high level of ancestral polymorphism in an out of equilibrium population due to high levels of clonal propagation and low levels of gene flow and recombination. Therefore, we assume that an extinction debt is present in C. dubium. As a management strategy, we propose to transplant ramets between multiple patches to increase the number of mating partners and therefore restore sexual reproduction.

Clonal Propagation of Valeriana jatamansi Retains the Essential Oil Profile of Mother Plants: An Approach Toward Generating Homogenous Grade of Essential Oil for Industrial Use

Valeriana jatamansi Jones (Syn. V. wallichii DC.) is an aromatic, medicinal herb used as a tranquilizer and in treating sleep disorders. Rhizome is mainly used to extract essential oil (EO) and valepotriates. High quality and economic yield of rhizomes are available in the third year of growth. Therefore, the cultivation of V. jatamansi is not picking up, and over-exploitation of this plant from wild habitats to meet the increasing demand of the pharmaceutical industry is the cause of threat to the genetic diversity of the species. Further, collections from the wild are heterogeneous, resulting in variable produce. The development of clonal lines can ensure uniform quality and yield of rhizome biomass. An effective clonal propagation method was standardized using different hormonal concentrations of naphthalene acetic acid (NAA) on apical shoot cuttings from the selected clone CSIR-IHBT-VJ-05 for different time durations and raised over various planting media. NAA treatment of 50 ppm concentration for 30 min was found optimum for root induction in apical shoots of V. jatamansi. Variations for EO composition within the clone were non-significant, while samples of the control population were variable. The best quality EO (patchouli alcohol ∼62%) was available during the third year of plant growth. A propagation technique for large-scale quality plant material (QPM) production has been standardized to reduce the stress over natural resources and promote V. jatamansi for use in the aromatic and pharmaceutical industry.

Clonal Propagation of Coffea arabica with Indole Butyric Acid and Acclimatization Conditions in Amazonas, Peru

The use of hormones is important in improving the propagation process. The objective of the experiment was to identify the best dose of indole butyric acid (IBA) hormone for the rooting of coffee cuttings of the typical variety through clonal propagation in the Amazon region. The experiment was carried out in two stages. Both stages were carried out under a Completely Randomized Design (CRD). The percentage of rooting of cuttings and the number of roots were evaluated. The coffee tree clones were collected from rust tolerant elite plants. In the laboratory, the cuttings were immersed for 10 minutes in fungicide and then in five doses of IBA hormone (1000, 1500, 2000, 2500, and 3000 mg L−1) of IBA. They were then placed in a microtunnel. With the best dose obtained in stage 1 of IBA (T4: 2000 mg L−1), which achieved a rooting percentage of 52.50% and 5.8 roots on average, stage 2 was installed for the evaluation of acclimatization conditions (greenhouse and nursery). Thirty plants of the Típica variety were evaluated per treatment. Regarding the results after 90 days, the greenhouse obtained the highest values with 72.22% survival, a plant height of 9.48 cm, 4.32 mm stem diameter, 6.53 leaves per plant, 9.65 cm leaf length, and 3.83 cm leaf width. The IBA hormone in a controlled environment such as the greenhouse achieves good vegetative propagation for the Típica variety.

Interhospital Transmission of Vancomycin-resistant Enterococcus Faecium in Aomori, Japan

A series of vancomycin-resistant Enterococcus faecium (VREf) outbreaks caused by clonal propagation due to inter-hospital transmission occurred in six general hospitals in Aomori prefecture, Japan. This was the first multi-jurisdictional outbreak of VREf sequence type 1421 in Japan. It took almost three years for VREf prevalence to return to pre-outbreak levels. The duration and size of outbreaks differed between hospitals according to the countermeasures available at each hospital. Prompt and strong countermeasures including patient isolation and repetitive screening of all inpatients were required to control the outbreaks. In addition to strict infection control measures, continuous monitoring of VREf prevalence in local medical regions and smooth and immediate communication among hospitals are required to prevent VREf outbreaks.

In Vitro Regeneration of Muscari racemosum Mill. Using Twin Bulb Scales, Primary Bulbs, and Leaf Bases

Turkey is an important center of diversity for many plants species including bulbs, rhizomes, tubers, and other plants of high agricultural and horticultural importance. These species have a special importance as ornamental plants. However, due to urbanization and related factors, many of them are under threat. One of these species is the endemic Muscari racemosum Mill. The current study aimed to develop an efficient in vitro commercial bulblet propagation procedure using different explants. Twin-scale bulb explants were regenerated on MS medium having several doses of Kinetin+NAA (1-Naphthaleneacetic acid). The best regeneration was exhibited on 4.65 μM Kinetin+5.37 μM NAA at the end of 10 weeks with induction of 4.08 bulblets/explant with a mean diameter of 0.31 cm. The primary bulblets were cultured on MS medium having 18.60 μM Kinetin+5.37 μM NAA. About a 2.5-fold increase in the diameter of the bulbs (0.76 cm) was exhibited on the regenerated bulblets. The bulblets were regenerated on leaf bases taken from MS medium having several doses of BAP (6-Benzylaminopurine) + NAA. The regenerated bulbs were rooted on MS medium having 4.90 μM IBA (Indole-3-butyric acid) followed by their transference to a greenhouse for acclimatization. This study provided important information on commercial clonal propagation of M. racemosum and the importance of explants and growth regulators in plant regeneration.