Isolation of endophytes from Dioscorea nipponica Makino for stimulating diosgenin production and plant growth.

KEY MESSAGE: Both bacterial and fungal endophytes exhibited one or more plant growth-promoting (PGP) traits. Among these strains, the Paenibacillus peoriae SYbr421 strain demonstrated the greatest activity in the direct biotransformation of tuber powder from D. nipponica into diosgenin. Endophytes play crucial roles in shaping active metabolites within plants, significantly influencing both the quality and yield of host plants. Dioscorea nipponica Makino accumulates abundant steroidal saponins, which can be hydrolyzed to produce diosgenin. However, our understanding of the associated endophytes and their contributions to plant growth and diosgenin production is limited. The present study aimed to assess the PGP ability and potential of diosgenin biotransformation by endophytes isolates associated with D. nipponica for the efficient improvement of plant growth and development of a clean and effective approach for producing the valuable drug diosgenin. Eighteen bacterial endophytes were classified into six genera through sequencing and phylogenetic analysis of the 16S rDNA gene. Similarly, 12 fungal endophytes were categorized into 5 genera based on sequencing and phylogenetic analysis of the ITS rDNA gene. Pure culture experiments revealed that 30 isolated endophytic strains exhibited one or more PGP traits, such as nitrogen fixation, phosphate solubilization, siderophore synthesis, and IAA production. One strain of endophytic bacteria, P. peoriae SYbr421, effectively directly biotransformed the saponin components in D. nipponica. Moreover, a high yield of diosgenin (3.50%) was obtained at an inoculum size of 4% after 6 days of fermentation. Thus, SYbr421 could be used for a cleaner and more eco-friendly diosgenin production process. In addition, based on the assessment of growth-promoting isolates and seed germination results, the strains SYbr421, SYfr1321, and SYfl221 were selected for greenhouse experiments. The results revealed that the inoculation of these promising isolates significantly increased the plant height and fresh weight of the leaves and roots compared to the control plants. These findings underscore the importance of preparing PGP bioinoculants from selected isolates as an additional option for sustainable diosgenin production.

Dendroclimatic response of Pinus tabuliformis Carr. along an altitudinal gradient in the warm temperate region of China.

Introduction: Global climate change can affect the sensitivity of tree radial growth to climate factors, but the specific responses of tree radial growth to microclimate along the altitudinal gradient in the long term are still unclear. Methods: In this study, the tree-ring width chronologies of Pinus tabuliformis Carr. in Shanxi Province of China were studied at three altitude gradients (1200-1300 m (low altitude), 1300-1400 m (medium altitude) and 1400-1500 m (high altitude)) during 1958-2017. Results: The results showed that (1) the climate background could be divided into two periods based on the Mann-Kendall test analysis: 1958-1996 was a stable period (mean annual temperature (MAT)=10.25°C, mean annual precipitation (MAP)=614.39 mm), and 1997-2017 was a rapid change period (MAT=10.91°C, MAP=564.70 mm), indicating a warming and drying trend in the study region. (2) The radial growth of P. tabuliformis at different altitudes showed inconsistent variation patterns. The tree radial growth at low and medium altitudes (CV=27.01% for low altitude and CV=24.69% for medium altitude) showed larger variation amplitudes during the rapid change period than that in the stable period (CV=12.40% for low altitude and CV=18.42% for medium altitude). In contrast to the increasing trend, the tree radial growth rates at the high altitude showed a decreasing trend across years. (3) In the stable period, the radial growth of P. tabuliformis at the low altitude showed a significantly negative response to temperature and a positive response to precipitation in May and June. The tree radial growth at the medium altitude was positively related to precipitation in June and minimum temperature in February. The tree growth at the high altitude was mainly positively correlated with the temperature in May and August. In the rapid change period, the radial growth of P. tabuliformis at the low altitude was affected by more meteorological factors than that in the stable period. Medium-altitude trees were positively influenced by precipitation in June and minimum temperature in January, whereas high-altitude trees responded positively to wind speed in February. (4) Along altitudinal gradients, tree radial growth was more related to temperature than precipitation in the stable period. The tree radial growth at the high altitude during the rapid change period was only affected by wind speed in February, whereas the tree radial growth at low and medium altitudes was mainly affected by temperature to a similar extent during the two periods. Discussion: The study indicated that tree growth-climate response models could help deeply understand the impact of climate change on tree growth adaptation and would be beneficial for developing sustainable management policies for forest ecosystems in the transition zone from warm-temperate to subtropical climates.

In vitro regeneration and its histological characteristics of Dioscorea nipponica Makino.

Dioscorea nipponica Makino is an optimal candidate to develop the diosgenin industry in North China. Due to its increasing demand in the medicine industry, it is urgent to apply new biotechnological tools to foster breeds with desirable traits and enhanced secondary metabolite production. The production of useful metabolites by the in vitro cultured rhizomes can be explored successfully for utilization by various food and drug industries. In this study, we reported callus formation and plantlet regeneration of the medicinal plant D. nipponica. Explants of leaves, stem segments and rhizomes of aseptic seedlings were cultured on Murashige and Skoog (MS) medium containing various combinations of auxin and cytokinin to find the optimal PGRs of each type of explant for callus induction and shoot regeneration of D. nipponica. The paraffin section technique was also used to observe of the morphogenesis of callus and adventitious bud. Explants of seeds and rhizomes formed calli at high frequency in all lines we examined. However, the explant of leaves rarely formed callus. Three kinds of callus were detected during the induction phase. Here, we describe three types of callus (Callus I-III) with different structure characteristics. Greenish in color and a nodule-like protrusion surface (Callus type III) were arranged more closely of cells with less interstitial substance, cell differentiation ability stronger than other callus types. The optimum combination was the maximum shoot differentiation frequency of 90% in callus derived from seeds cultured on MS medium with 2.0 mg L-16-BA + 0.2 mg L-1NAA. The shoot differentiation frequency (88.57%) of rhizome-induced callus was obtained by the combination of MS medium supplemented with 3.0 mg L-16-BA + 2.0 mg L-1NAA. 1/2 MS medium plus 0.5 mg L-1NAA resulted in a higher root regeneration frequency of 86.70%. In vitro propagated plantlets with healthy roots were domesticated and transplanted into small plastic pots containing sterile soil rite under greenhouse conditions with 80% survivability. Bud differentiation is mostly of exogenous origin, mostly occurring on the near callus surface. Therefore, it may be surmised that in vitro morphogenesis of D. nipponica is mainly caused by indirect organogenesis (adventitious bud).