Leveraging machine learning to unravel the impact of cadmium stress on goji berry micropropagation.

This study investigates the influence of cadmium (Cd) stress on the micropropagation of Goji Berry (Lycium barbarum L.) across three distinct genotypes (ERU, NQ1, NQ7), employing an array of machine learning (ML) algorithms, including Multilayer Perceptron (MLP), Support Vector Machines (SVM), Random Forest (RF), Gaussian Process (GP), and Extreme Gradient Boosting (XGBoost). The primary motivation is to elucidate genotype-specific responses to Cd stress, which poses significant challenges to agricultural productivity and food safety due to its toxicity. By analyzing the impacts of varying Cd concentrations on plant growth parameters such as proliferation, shoot and root lengths, and root numbers, we aim to develop predictive models that can optimize plant growth under adverse conditions. The ML models revealed complex relationships between Cd exposure and plant physiological changes, with MLP and RF models showing remarkable prediction accuracy (R2 values up to 0.98). Our findings contribute to understanding plant responses to heavy metal stress and offer practical applications in mitigating such stress in plants, demonstrating the potential of ML approaches in advancing plant tissue culture research and sustainable agricultural practices.

Lycibarbarines A-C, Three Tetrahydroquinoline Alkaloids Possessing a Spiro-Heterocycle Moiety from the Fruits of Lycium barbarum.

Three tetrahydroquinoline alkaloids, lycibarbarines A-C (1-3), possessing a unique tetracyclic tetrahydroquinoline-oxazine-ketohexoside fused motif, were isolated from the fruits of Lycium barbarum. Their structures were determined by spectroscopic analysis and quantum-chemical calculations. Compounds 1 and 3 exhibited neuroprotective activity when evaluated for corticosterone-induced injury by reducing the apoptosis of PC12 cells through the inhibition of caspase-3 and caspase-9.

Semen quality improvement in boars fed with supplemental wolfberry (Lycium barbarum).

Wolfberry is well known for its health benefits in Asian countries. This study consisted of two experiments. In Experiment 1, nine boars were provided 40 g dried wolfberry per 100 kg body weight per day in addition to regular feed for 160 days (divided into 40 days phases: I, II, III, and IV) under step-down air temperature conditions. Controls (n = 9) were fed regular feed only. Significant (p < .05 or p < .01) or slight improvements in sperm progressive motility, total abnormality rate, sperm concentration, and total sperm per ejaculate were observed in the wolfberry group during phases II and III. No differences were observed in semen volume. After combining the data from phases II ~ IV, significant improvements were detected in all aforementioned traits (p < .05 or p < .01), except semen volume. In Experiment 2, the wolfberry group (n = 5) was fed wolfberry for 90 days and exhibited significantly reduced head, tail, and total abnormality rates (p < .05 or p < .01) in both fresh semen and semen stored for 72 hr at 17°C compared to the control group (n = 5). SOD activity also significantly increased in this group of boars. Collectively, the findings of this study suggest that wolfberry has a positive effect on boar semen quality.

Impact of Nitrogen Fertilizer Levels on Metabolite Profiling of the Lycium barbarum L. Fruit.

The yield and quality of goji (Lycium barbarum L.) fruit are heavily dependent on fertilizer, especially the availability of nitrogen, phosphorus, and potassium (N, P, and K, respectively). In this study, we performed a metabolomic analysis of the response of goji berry to nitrogen fertilizer levels using an Ultra Performance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry (UPLC-ESI-MS/MS) method. There was no significant difference in the fruit yield or the commodity grade between N0 (42.5 g/plant), N1 (85 g/plant), and N2 (127.5 g/plant). The primary nutrients of the goji berry changed with an increasing nitrogen fertilization. Comparative metabolomic profiling of three nitrogen levels resulted in the identification of 612 metabolites, including amino acids, flavonoids, carbohydrates, organic acids, and lipids/alcohols, among others, of which 53 metabolites (lipids, fatty acids, organic acids, and phenolamides) demonstrated significant changes. These results provide new insights into the molecular mechanisms of the relationship between yield and quality of goji berry and nitrogen fertilizer.

Phytotoxic effects of bulk and nano-sized Ni on Lycium barbarum L. grown in vitro - Oxidative damage and antioxidant response.

This study aimed to evaluate the effects of nickel oxide nanomaterial (nano-NiO) on goji berry (Lycium barbarum L.) shoots grown under in vitro conditions and to determine if the nanomaterial was more harmful than its bulk counterpart, nickel (II) sulphate (NiSO4). For this purpose, in vitro shoots of L. barbarum were cultured on MS medium supplemented with 15 mg L-1 of NiSO4 or nano-NiO. Nano-NiO was more harmful for shoots growth and photosynthetic pigments than NiSO4, with reductions up to 82% in comparison to the control. Shoots treated with nano-NiO presented an overproduction of hydrogen peroxide (H2O2; 130% increase) and superoxide anion (O2-; 110% increase), which led to higher levels of lipid peroxidation (LP; 57% increase) and the occurrence of oxidative stress. In opposition, bulk Ni seemed not to induce oxidative stress, once LP and reactive oxygen species content decreased in comparison with the control. The evaluation of the non-enzymatic antioxidant (AOX) system revealed that, under nano-NiO excess, proline, ascorbate, glutathione and phenols levels increased up to 4-fold, but did not change in response to the bulk treatment. With respect to the enzymatic AOX system, nano-NiO enhanced the activities of superoxide dismutase (203%) and ascorbate peroxidase (62%), though catalase activity was negatively affected, while bulk Ni did not majorly affect these enzymes' behavior. Overall, the data showed that Ni phytotoxicity in L. barbarum shoots depends on the metal source and that, in this case, nano-NiO seemed to be more deleterious to goji shoots grown under in vitro conditions than NiSO4.

[Study on membrane type leaf water evaporation inhibitors for improving effect of preventing diseases and pest controlling of Lycium barbarum].

Through indoor and field comparative experiments, the properties of membrane type leaf evaporation inhibitors and its effects on photosynthesis of Lycium barbarum and compatibility and synergistic of pesticide were studied. The evaporation inhibitors and L. barbarum were chosen to investigate the suppression of water evaporation and the compatibility with pesticides. The effect of evaporation inhibitors on photosynthesis of L. barbarum leaves was determined by the chlorophyll fluorescence imaging system. The results showed that water evaporation of L. barbarum leaves of different leaf age were evidently suppressed after treated with evaporation inhibitor. The inhibitor was well compatible with pesticide and effectively improved the pesticide efficacy，and had no significant effect on chlorophyll fluorescence parameters. It is concluded that the evaporation inhibitor has good compatibility with the pesticide, and has remarkable effect of restraining moisture evaporation, which make it can be used for reducing the dosage and improving the efficacy of the pesticide in the field of L. barbarum.

Influence of Rhizoglomus irregulare on nutraceutical quality and regeneration of Lycium barbarum leaves under salt stress.

Whether arbuscular mycorrhizal fungi augment the nutraceutical quality of crops under salt stress is critical as a potential agronomic practice in salinized farmland. To evaluate the effect of Rhizoglomus irregulare on the nutraceutical quality of Lycium barbarum leaves under salt stress, we analyzed growth parameters and the rutin, polysaccharide, acidic polysaccharide, and amino acids contents of 2 harvests. Inoculation of R. irregulare significantly increased the regenerated bud number (partial eta squared (PES) = 0.577, P < 0.0001) and rutin concentration (PES = 0.544, P < 0.001) of L. barbarum leaves, with and without salt stress. The biomass of the 2nd harvest (PES = 0.355, P = 0.0091) and acidic polysaccharide (PES = 0.518, P = 0.001) of L. barbarum leaves were notably enhanced by R. irregulare under 200 mmol/L salt level. Rhizoglomus irregulare had insignificant effect on polysaccharide (PES = 0.092, P = 0.221) and amino acids levels (PES = 0.263, P = 0.130) in the leaves of L. barbarum. However, inoculation by R. irregulare decreased proline level (PES = 0.761, P = 0.001) in the leaves of L. barbarum when subjected to salt stress. Taken together, these results indicate that R. irregulare significantly improved the nutraceutical quality and facilitated the sustainable production of L. barbarum leaves exposed to salt stress.

The effects of exogenous antioxidant germanium (Ge) on seed germination and growth of Lycium ruthenicum Murr subjected to NaCl stress.

In this paper, we present the results of a study on the effects of exogenous antioxidant germanium (Ge) on seed germination and seedling growth, and its role as a radical scavenger that regulates related enzymes, including superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT), under salt stress. Seeds were incubated in 0, 50, 100, 150, 200, 250 and 300 mM NaCl to determine the salt tolerance of the Lycium ruthenicum Murr seedlings and from the results, the critical and ultimate salt concentrations were chosen for the next experiment. Subsequently, two treatments (seeds soaked in Ge and Ge added to salt) with four concentrations of GeO2 (0, 5, 10 and 20 µM) were used with the critical (150 mM) and ultimate salt concentrations (250 mM). The results demonstrated that salt alone inhibited seed germination significantly (≥150 mM) and reduced seedling growth (≥200 mM). The addition of exogenous Ge to the salt solution, as well as soaking the seeds in Ge, attenuated the salt stress effects in a manner dependent on the dose of Ge, as indicated by the increased percentage of seeds that germinated and improved seedling growth. The addition of Ge also showed a significant reversal of salt stress on the activities of antioxidant enzymes, with a decrease in SOD and POD activity, but an increase in CAT activity with 150 mM NaCl, and enhancement of SOD, POD and CAT with 250 mM NaCl. Correspondingly, the level of malondialdehyde was decreased significantly by each Ge treatment under salt stress. Further, for L. ruthenicum, adding 10 Ge and seeds soaked in 5 Ge were the most effective treatments. To our knowledge, this is the first report to show the protective effects of exogenous Ge against salt-induced oxidative damage in L. ruthenicum seed germination and seedling growth. Thus, L. ruthenicum can be used in areas with salty soil and Ge can promote the plants' salt tolerance.

Identification and validation of reference genes for quantitative real-time PCR normalization and its applications in lycium.

Lycium barbarum and L. ruthenicum are extensively used as traditional Chinese medicinal plants. Next generation sequencing technology provides a powerful tool for analyzing transcriptomic profiles of gene expression in non-model species. Such gene expression can then be confirmed with quantitative real-time polymerase chain reaction (qRT-PCR). Therefore, use of systematically identified suitable reference genes is a prerequisite for obtaining reliable gene expression data. Here, we calculated the expression stability of 18 candidate reference genes across samples from different tissues and grown under salt stress using geNorm and NormFinder procedures. The geNorm-determined rank of reference genes was similar to those defined by NormFinder with some differences. Both procedures confirmed that the single most stable reference gene was ACNTIN1 for L. barbarum fruits, H2B1 for L. barbarum roots, and EF1α for L. ruthenicum fruits. PGK3, H2B2, and PGK3 were identified as the best stable reference genes for salt-treated L. ruthenicum leaves, roots, and stems, respectively. H2B1 and GAPDH1+PGK1 for L. ruthenicum and SAMDC2+H2B1 for L. barbarum were the best single and/or combined reference genes across all samples. Finally, expression of salt-responsive gene NAC, fruit ripening candidate gene LrPG, and anthocyanin genes were investigated to confirm the validity of the selected reference genes. Suitable reference genes identified in this study provide a foundation for accurately assessing gene expression and further better understanding of novel gene function to elucidate molecular mechanisms behind particular biological/physiological processes in Lycium.

[Effect of exogenous calcium on seed germination and seedling physiological characteristics of Lycium ruthenium].

OBJECTIVE: In order to get the method for improving the salt resistance of Lycium ruthenium seeds and seedlings under NaCl stress, the seed germination and physiological characteristics of L. ruthenium seedlings was studied. METHOD: Several physiological indexes of L. ruthenium seeds under NaCl stress, such as the germination rate (Gr), germination vigor (Gv), germination index (Gi), vigor index (Vi), and relative salt damage rate were measured. Other indexes of the seedlings like relative water contents (RWC) , chlorophyll contents, soluble protein contents, electrolyte leakage, the contents of malondialdehyde (MDA), and peroxidase (POD) were also measured. RESULT: NaCl at lower concentration could promote the seed germination but inhibit the seed germination at higher concentration. After the treatment by CaCl2 at the different concentrations, all germination indexes were increased. With the increase of salt concentration, the relative water contents and the contents of chlorophyll were decreased, the content of MDA and electrolyte leakage were increased. The change trend of POD activity showed the first increase and then decrease with the increase of salt concentration, which was similar to that of the soluble protein. After the treatment by CaCl2, relative water contents, chlorophyll and POD activities were decreased more slowly, and also electrolyte leakage and MDA contents increased slowly. CONCLUSION: The CaCl2 could significantly alleviate the damages to the seeds and seedlings of L. ruthenium under NaCl stress, and promote the salt resistance to the seeds and seedlings of L. ruthenium.

[Controling of Paratrioza sinica by bionic glue].

OBJECTIVE: To investigate the controlling effects of bionic glue on Paratrioza sinica. METHOD: P. sinica and bionic glue were chosen as materials to investigate the adhesive rate, mortality rate, and study the effects of behavior of P. sinica and growth of leaves sprayed with bionic glue. RESULT: Spraying of the bionic glue can significantly increase the adhesive rate of P. sinica with no obviously repellency, and it can be used to control the adults of P sinaca in field with no significant effect on the growth of wolfberry leaves. CONCLUSION: Bionic glue has significant controlling effects on adults of P. sinica, it can be used as an eliminator of adults of P. sinica in field at the beginning of the vegetation season to suppress of the development of P. sinica population.

Microcutting culture and morpho-physiological changes during acclimation in two Lycium chilense cytotypes

Lycium chilense, a deciduous perennial shrub, is one of the endangered native species of Patagonia due to sheep overgrazing. Chances of recolonization by seeds are scarce due to the limited density of propagules in the soil and very specific requirements for germination. The objective was to develop an in vitro propagation protocol that would help to perform reestablishment of this species in degraded areas of the Patagonian steppe. Seeds came from two provenances with different somatic number due to differences in ploidy level. Defoliated microcuttings were planted in test tubes with different growing media and taken to a growth chamber. Rooting percentage did not differ between origins, but higher values were encountered for medium without hormones. Subcultures increased significantly rooting percentage and reduced time to rooting. The leaves from micropropagated plants were thinner, did not exhibit hairs, and had poorly developed palisade parenchyma and less epicuticular waxes. In vitro leaves had lower stomatal density and their stomata were less functional when compared to acclimated leaves. A repopulation program of Lycium chilense based on microcutting culture, specialy using subcultures, is feasible. (AU)

Microcutting culture and morpho-physiological changes during acclimation in two Lycium chilense cytotypes.

Lycium chilense, a deciduous perennial shrub, is one of the endangered native species of Patagonia due to sheep overgrazing. Chances of recolonization by seeds are scarce due to the limited density of propagules in the soil and very specific requirements for germination. The objective was to develop an in vitro propagation protocol that would help to perform reestablishment of this species in degraded areas of the Patagonian steppe. Seeds came from two provenances with different somatic number due to differences in ploidy level. Defoliated microcuttings were planted in test tubes with different growing media and taken to a growth chamber. Rooting percentage did not differ between origins, but higher values were encountered for medium without hormones. Subcultures increased significantly rooting percentage and reduced time to rooting. The leaves from micropropagated plants were thinner, did not exhibit hairs, and had poorly developed palisade parenchyma and less epicuticular waxes. In vitro leaves had lower stomatal density and their stomata were less functional when compared to acclimated leaves. A repopulation program of Lycium chilense based on microcutting culture, specially using subcultures, is feasible.

Effects of polyamines on organogenesis and somatic embryogenesis of Lycium barbarum calli.

Levels of three endogenous free polyamines (PAs), Put, Spd and Spm, were detected during organogenesis and somatic embryogenesis (SE) of Lycium barabrum (L.) calli. The predominant forms of PAs in organogenesis and SE were found to be Put and Spd, respectively. In both developmental pathways, the changes of Put content were very similar, i.e., it accumulated quickly in the initial stages of calli differentiation and then decreased; with the further morphogenesis, increase in Put level was also observed. The highest level of Spd was obtained at day 1 of calli organogenesis; while, after 1 day of culture for calli SE, Spd level began to increase and reached a maximum at day 10. The treatments with exogenous PAs had positive effects on both organogenesis and SE, especially those of Spd on organogenesis and SE and Put on SE. However, Put showed no effects on adventitious bud formation. CHA, which inhibits the activity of Spd synthetase, prevented both adventitious bud formation and further development of somatic embryo into plantlet. Although MGBG, a specific inhibitor of S-adenosyl methionine decarboxylases, had little effects on organogenesis, it reduced the number of somatic embryo and the plantlets subsequently regenerated. Such inhibitions could be reversed by Spd (50 mumol/L). These results from organogenesis and SE of the same specie indicate that PAs influenced these two in vitro morphogenesis pathways.