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An efficient protocol for regenerating shoots from paper mulberry (Broussonetia papyrifera) leaf explants

  • Siming Cui , Ying Ren , Yahan Hao , Junjie Zhang , Zhouchao Chen , Jintuo Zou , Wei Zhou EMAIL logo and Xiaoyang Chen EMAIL logo
Published/Copyright: June 10, 2020
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Open Life Sciences
From the journal Open Life Sciences Volume 15 Issue 1

Abstract

Paper mulberry (Broussonetia papyrifera) is a tree species that has many economic, ecological, and social uses. This study developed an efficient protocol for regenerating shoots from leaf explants using Murashige and Skoog (MS) medium supplemented with different concentrations of plant growth regulators (PGRs), which play vital roles in shoot regeneration. The best result, 86.67% induction frequency and 4.35 shoots per explant, was obtained in the MS medium containing 2.0 mg/L N6-benzyladenine (BA) and 0.05 mg/L indole-3-butyric acid. The effects of explant age, orientation, and genotype were also investigated. Explants from young leaves had a greater regeneration frequency than those from old leaves, and the results were better when the distal end of the leaf explant contacted the medium versus the proximal end. Approximately 70.96% of the shoots rooted well in the MS medium containing 0.4 mg/L α-naphthalene acetic acid (NAA). Although some genotypes achieved poorer results, the regeneration protocol is still applicable for mass multiplication and genetic transformation.

Keywords: paper mulberry; leaf explant; plant regeneration; explant age; inoculation orientation

1 Introduction

Paper mulberry (Broussonetia papyrifera) is a multipurpose tree or shrub belonging to the Moraceae [1] that mainly grows in eastern Asia and the Pacific islands [2,3,4]. Each organ of the tree species is valuable: bark fiber is an excellent raw material for paper-making and a potential bio-ethanol feedstock [5,6,7] and young shoots have a high feed and nutritional value [8]. There are many medicinal uses for paper mulberry as a source of flavonoids, terpenoids, polyphenols, alkaloids, and other compounds [9,10,11,12]. In summary, paper mulberry has ecological, economic, forage, and medicinal values and has good development prospects. Consequently, paper mulberry is attracting increasing attention worldwide. Paper mulberry has strong stress resistance and environmental adaptability [13,14]; it is drought resistant and has the strong anti-pollution ability [15,16], allowing it to play an important role in ecological restoration [17,18,19].

The standard method of paper mulberry proliferation is seed breeding, but the survival rate is low and uniform seeding is not obtained by seed proliferation [20]. Paper mulberry can also proliferate through root cultivation and cutting propagation [21,22], but the efficiency is relatively low and cannot meet large-scale production needs. Therefore, to meet the demand for large-scale production and achieve efficient, convenient propagation, it is important to establish an efficient regeneration protocol for paper mulberry using a genetic transformation system based on the fundamental molecular and biochemical characteristics of this plant [23]. Several B. papyrifera regeneration protocols have been reported: proliferation from stem explants cultured in various plant growth regulator (PGR) combinations [24,25]; shoot regeneration from hypocotyl explants cultured on Murashige and Skoog (MS) medium containing 0.5–1.5 mg/L N6-benzyladenine (BA) and 0.1–0.5 mg/L naphthalene acetic acid (NAA) [26,27]; and shoot proliferation from leaf explants cultured on the MS medium supplemented with 1.5–2.0 mg/L BA and 0.05–1.0 mg/L NAA [28,29,30,31]. However, the proliferation efficiency of these systems is low and not satisfactory for establishing an efficient regeneration protocol. The selection of an effective explant is a prerequisite for any plant regeneration protocol [32]. Leaves are abundant and allow for stable gene transfer, but there is no efficient regeneration protocol using leaves as explants. This study explored the effects of PGRs, age, orientation, and genotype on the regeneration efficiency to establish an efficient shoot regeneration protocol from paper mulberry leaf explants for application in mass multiplication and genetic transformation.

2 Materials and methods

2.1 Plant materials, explant sources, and growth conditions

Seeds were collected from the Guangzhou cultivar in a field at South China Agricultural University (genotypes not included). Seeds were soaked in concentrated sulfuric acid for 9 min and washed with tap water for 5 min to soften the seed coat. After sterilization with 75% (v/v) ethanol for 50 s and 25% (w/v) NaClO for 15 min, the seeds were rinsed four times with sterile water and inoculated onto the MS medium [26]. When the seeds grew to seedlings after 4 weeks, seedling leaves were used as explants for experiments. All cultures were kept under cool white light (approximately 50 µmol/m2/s) with a 12 h photoperiod at 24 ± 1°C.

2.2 Induction and growth of shoot-buds

To compare the PGRs influencing the culture response, BA (0.0, 1.0, 1.5, 2.0, and 2.5 mg/L), indole-3-butyric acid (IBA; 0.0, 0.01, 0.05, and 0.10 mg/L), and kinetin (KT; 0.0, 0.5, and 1.0 mg/L) were used in combination in the MS basal medium. To investigate the effect of explant age on shoot regeneration, seedlings were divided into three sections from top to bottom: Section 1, top 1–3 leaves; Section 2, 4–6 leaves in the middle; and Section 3, bottom 7–9 leaves. To investigate the effect of explant orientation on shoot regeneration, we performed an experiment to examine the effects of contact of the proximal and distal ends with the medium. Four tree cultivars were used to examine the influence of genotype on shoot regeneration: NC-1, NC-2, TS-1, and ZQ-1. The percentage induction of shoot-buds and the number of shoot-buds per explant were recorded after 5 weeks of culture.

2.3 Rooting and acclimatization

The shoots were cut from the tissue and transferred onto the MS basal medium supplemented with NAA (0.0, 0.1, 0.2, 0.4, and 0.8 mg/L) to facilitate root initiation and the growth of intact plantlets. Plantlets with well-developed roots were removed from the culture bottles and washed under running tap water to remove any remaining medium. Then, the plantlets were placed in plastic pots filled with planting substrate in a greenhouse at high humidity. After acclimatization for 3 weeks, the plantlets were ready for transplantation in the field.

2.4 Data collection and statistical analysis

The experiments were conducted in a randomized complete block design; each treatment consisted of ten explants and was repeated three times. Analysis of variance (ANOVA) was carried out using SPSS 22.0. Duncan’s multiple range test was used to detect differences among means, with a significance level of 0.05.

3 Results and discussion

3.1 Effect of PGRs on shoot-bud induction

The seedlings (Figure 1a) were removed from the culture bottles aseptically, and only the leaves from Section 1 were used as explants for the experiment. The leaves were excised, cut into 5 × 5 mm2 pieces, and inoculated on the MS medium supplemented with various concentrations and combinations of BA, IBA, and KT for shoot induction. In the in vitro regeneration process, leaves began to expand gradually, the thickness gradually increased, the color changed from green to light, and the leaves were wrinkled and curled in 1 week. Shoot-buds appeared at 3 weeks (Figure 1b). Only parts of the induced shoot-buds elongated to form shoots.

Figure 1 Shoot regeneration from paper mulberry leaf explants and acclimatization of the regenerated plantlets. (a) 2 week-old seedling; (b) numerous shoot buds developed from leaf explants in the MS medium with 2.0 mg/L BA and 0.05 mg/L IBA in 3 weeks; (c) 2–3 cm-high shoots recovered from leaf explants in the MS medium with 2.0 mg/L BA and 0.05 mg/L IBA for 4 weeks; (d) root systems of regenerated shoot buds in the MS medium supplemented with 0.4 mg/L NAA; (e) an intact regenerated plantlet; and (f) an acclimatized plant transferred to a plastic pot filled with planting substrate.
Figure 1

Shoot regeneration from paper mulberry leaf explants and acclimatization of the regenerated plantlets. (a) 2 week-old seedling; (b) numerous shoot buds developed from leaf explants in the MS medium with 2.0 mg/L BA and 0.05 mg/L IBA in 3 weeks; (c) 2–3 cm-high shoots recovered from leaf explants in the MS medium with 2.0 mg/L BA and 0.05 mg/L IBA for 4 weeks; (d) root systems of regenerated shoot buds in the MS medium supplemented with 0.4 mg/L NAA; (e) an intact regenerated plantlet; and (f) an acclimatized plant transferred to a plastic pot filled with planting substrate.

The experimental results are shown in Tables 1–3. It is clear that PGRs played a vital role in shoot regeneration, and the proliferation at different concentrations was much better than when the PGRs were used alone (Tables 1 and 2). Although BA was more effective at promoting paper mulberry shoot regeneration than IBA, the best results were obtained when they were used together (Tables 1 and 2). KT had an inhibitory effect; the best results were obtained without KT (Table 3). The greatest efficiency was achieved when the MS basal medium was supplemented with 2.0 mg/L BA and 0.05 mg/L IBA: the shoot induction efficiency was 80.33% and there were 4.33 regenerated shoots (Tables 1–3).

Table 1

Effect of different BA concentrations on shoot-bud induction

BA (mg/L)Explants that regenerated shoots (%)Shoots per explant
0.00 e0 e
1.023.33 ± 13.02 cd2.33 ± 0.26 cd
1.550.00 ± 10.86 ab3.70 ± 0.34 ab
2.080.33 ± 11.68 a4.33 ± 0.56 a
2.536.67 ± 12.51 bc3.40 ± 0.42 bc

Each value represents the mean ± standard error of three replicates, each with 10 explants. The medium contained 0.05 mg/L IBA and 0.0 mg/L KT. Means followed by the same letter in the same column are not significantly different from each other at P ≤ 0.05, according to Duncan’s multiple range test.

Table 2

Effect of different IBA concentrations on shoot-bud induction

IBA (mg/L)Explants that regenerated shoots (%)Shoots per explant
0.00 13.33 ± 9.08 b2.73 ± 0.36 b
0.01 56.67 ± 13.33 ab3.57 ± 0.41 ab
0.05 80.33 ± 11.68 a4.33 ± 0.46 a
0.10 10.00 ± 8.51 c2.87 ± 0.26 c

Each value represents the mean ± standard error of three replicates, each with 10 explants. The medium contained 2.0 mg/L BA and 0.0 mg/L KT. Means followed by the same letter in the same column are not significantly different from each other at P ≤ 0.05, according to Duncan’s multiple range test.

Table 3

Effect of different KT concentrations on shoot-bud induction

KT (mg/L)Explants that regenerated shoots (%)Shoots per explant
0.0 80.33 ± 11.68 a4.33 ± 0.56 a
0.5 46.33 ± 12.98 b2.93 ± 0.34 b
1.0 26.67 ± 11.01 b1.87 ± 0.25 b

Each value represents the mean ± standard error of three replicates, each with 10 explants. The medium contained 2.0 mg/L BA and 0.05 mg/L IBA. Means followed by the same letter in the same column are not significantly different from each other at P ≤ 0.05, according to Duncan’s multiple range test.

Several regeneration protocols using leaves for explants have been reported for B. papyrifera. For example, Huang et al. [31] found that culturing paper mulberry leaf explants on the MS medium supplemented with 2.0 mg/L BA and 0.1 mg/L NAA had the best induction efficiency (81.7%), the regeneration efficiency was similar to this experiment, but the results did not show the number of shoots per explant, which was not rigorous. Liu et al. [30] found that regenerating shoots with leaf explants on the MS medium containing 1.5 mg/L BA and 1.0 mg/L NAA created the maximum number of shoots (3.88 shoots per explant), which was less than this experiment, and the experimental results did not show the regeneration efficiency, which was imperfect. However, Yu et al. [28] achieved a shoot induction efficiency of 36.4% propagating paper mulberry using leaf explants only, which was low. The effects of BA in organogenesis are supported by similar reports in other plants [33,34]. These results suggest that cell activities during shoot-bud regeneration are controlled by various internal factors including cytokinins, which may cause internal chemical and structural differences, leading to different responses when present in the medium. It is worth mentioning that NAA was used for auxin in the above studies, and IBA was used in this study. Obviously, the results of this study are better, and similar results have been reported in hemp [35]. However, the specific roles of BA and IBA remain unclear [26].

3.2 Effect of explant age on shoot-bud regeneration

Explant age affected the frequency of shoot-bud regeneration and the mean number of shoots per explant (Table 4). Explants from Section 1 had the greatest regeneration frequency (86.67%) and produced 4.35 shoot buds per explant. Therefore, the explant age plays an important role in shoot regeneration. Similar results were obtained in other plants. For example, Vaidya et al. [36] found that the age of peppermint shoot tip explants affected regeneration frequency; Singh [37] observed that the regeneration efficiency of Jatropha curcas was affected by the age of leaf explants; explants from younger seedlings (≤15 days) were still juvenile and formed callus easily, whereas beyond 30 days, the regeneration response decreased with an increase in the age of seedlings. Younger explants have greater morphogenic potential than older explants, as they might have more metabolically active cells due to hormonal, nutritional, and other physiological conditions that are responsible for increased organogenesis [33]. However, Zhang et al. [23] found that for Moringa, the explants were either too young or too old with unsatisfactory results. This may be due to the different meristematic capabilities of different plants. Each plant only has enough metabolically active cells if it grows to a specific age.

Table 4

Effect of explant age on shoot regeneration and elongation

Leaf age Explants that regenerated shoots (%)Shoots per explant
Section 186.67 ± 12.33 a4.35 ± 0.22 a
Section 276.67 ± 13.02 a3.23 ± 0.56 ab
Section 360.00 ± 11.82 ab2.53 ± 0.31 ab

Each value represents the mean ± standard error of three replicates, each with 10 explants. The medium contained 2.0 mg/L BA, 0.05 mg/L IBA, and 0.0 mg/L KT. Means followed by the same letter in the same column do not differ significantly from each other at the P ≤ 0.05 level, according to Duncan’s multiple range test.

3.3 Effect of explant orientation on shoot-bud propagation

The orientation of the explants significantly influenced the shoot-bud induction response. When the distal end of the leaf explants contacted the medium, the shoot-bud regeneration rate reached 73.33%. However, this rate decreased to 13.67% when the proximal end of the explant contacted the medium. The number of induced shoot buds per explant increased from 1.02 to 3.58 after placing the distal end of the leaf explants contacted the medium (Table 5). In the cultures, shoot buds were mostly induced from the vein and edge of the cut surface of the distal end, as has been reported for sugarcane [38]. Similarly, Kumar et al. [39] found that Salvadora oleoides had a higher regeneration efficiency when the distal end of leaf explants contacted the medium. When studying the effect of explant orientation on regeneration efficiency, not only leaf explants, Liu et al. [40] found that for petiole explants of Jatropha curcas L., the method of inoculating the explants horizontally on the medium was more beneficial to the regeneration than inoculating the explants vertically. These results may be related to the transport of polar auxin, because the intensity of auxin polar transport is inversely related to the sensitivity of rooting [41]. Moreover, the logical explanation for the effects of orientation of the explants may be the oxygen tension in the meristematic cells at the explant–medium interface [38]. However, the specific reasons are not clear, and further research is needed.

Table 5

Effect of explant orientation on shoot propagation and elongation

OrientationExplants that regenerated shoots (%)Shoots per explant
Distal end 73.33 ± 14.983.58 ± 0.42
Proximal end 13.67 ± 9.911.02 ± 0.27

Each value represents the mean ± standard error of three replicates, each with 10 explants. The medium contained 2.0 mg/L BA, 0.05 mg/L IBA, and 0.0 mg/L KT. Means followed by the same letter in the same column do not differ significantly from each other at the P ≤ 0.05 level, according to Duncan’s multiple range test.

3.4 Effect of explant genotype on shoot-bud regeneration

To examine the adaptability of the regeneration protocol to different paper mulberry trees, four cultivars with different genetic backgrounds were tested and their regeneration efficiencies and numbers of shoot-buds per explant were compared. As shown in Table 6, the regeneration efficiency and number of shoot-buds per explant varied greatly among the genotypes grown under the same conditions. Explants from ZQ-1 and NC-2 had higher regeneration frequencies of 83.33% and 78.58%, respectively, while explants from NC-1 and TS-1 fared much worse, with regeneration frequencies of 56.67% and 46.67%, respectively. The reason for this difference may be that the endogenous hormone levels varied amongst calli derived from different genotypes and explant types [42]. The differential behavior can be related to different mechanisms for control of the endogenous PGRs metabolism and/or contents. The regeneration capacity of calli is strongly correlated with the concentration of endogenous hormone; recently, Kudo et al. [43] suggested that the endogenous hormone levels affect the regeneration ability of calli. Similar results have been reported in other plants, including Mucuna bracteata DC. ex Kurz [44], Punica granatum L. [45], conifers [46], J. curcas [47], and jojoba [48].

Table 6

Effect of explant genotype on shoot regeneration and elongation

GenotypeExplants that regenerated shoots (%)Shoots per explant
NC-156.67 ± 13.33 b3.57 ± 0.41 a
NC-278.58 ± 10.69 a3.88 ± 0.27 a
TS-146.67 ± 9.88 b2.33 ± 0.32 b
ZQ-183.33 ± 12.33 a4.21 ± 0.38 a

Each value represents the mean ± standard error of three replicates, each with 10 explants. The medium contained 2.0 mg/L BA, 0.05 mg/L IBA, and 0.0 mg/L KT. Means followed by the same letter in the same column do not differ significantly from each other at the P ≤ 0.05 level, according to Duncan’s multiple range test.

3.5 Rooting and acclimatization

Shoots 2–3 cm in height (Figure 1c) were placed on rooting medium after 4 weeks. The percentage of rooting and the number and length of roots of each seedling increased with the concentration of NAA added to the MS basal medium (Table 7). The greatest rooting percentage (Figure 1d and e) achieved was 70.96% when shoots were placed on the MS medium containing 0.4 mg/L NAA. However, roots induced on the MS medium with higher concentrations of NAA (0.8 mg/L) developed more, but shorter, weaker roots. A similar observation has been observed in Simmondsia chinensis [49].

Table 7

Effect of NAA on root induction and growth

NAA (mg/L)Rooting (%)Root no.Root length (cm)
0.010.33 ± 8.25 c3.56 ± 0.42 c0.71 ± 0.15 b
0.150.28 ± 12.33 b2.82 ± 0.25 c1.13 ± 0.18 ab
0.270.23 ± 11.28 a5.33 ± 1.58 bc1.28 ± 0.23 a
0.470.96 ± 10.56 a10.68 ± 1.33 b1.75 ± 0.31 a
0.850.34 ± 12.48 b16.53 ± 1.54 a0.73 ± 0.18 b

Each value represents the mean ± standard error of three replicates, each with 10 explants. Means followed by the same letter in the same column do not differ significantly from each other at the P ≤ 0.05 level, according to Duncan’s multiple range test.

The rooted shoots survived well after transfer to plastic pots filled with planting substrate (Figure 1f). The acclimatization of the rooted shoots was accomplished and more than 90% of the plants were successfully transferred to plastic pots in greenhouse conditions. The results indicated that the regenerated shoots had good environmental adaptability. Similar results have also been obtained in other plants. Cheng et al. [35] found that 75% of the hemp-rooted shoots survived after acclimation. Similarly, Liu et al. [40] observed that more than 80% of J. curcas were successfully transferred to plastic pots. These results suggest that most of the regenerated seedlings can survive well after transfer to plastic pots, but the survival rate of different plants is different, probably because their environmental adaptability is different.

4 Conclusions

An efficient shoot regeneration protocol from B. papyrifera leaf explants was established. The greatest shoot induction efficiency was 86.67%, and the maximum number of shoots regenerated was 4.35. The plants produced with this protocol were healthy and had good environmental adaptability. This method will be useful for subsequent application in mass multiplication and genetic transformation.

Acknowledgments

This research was funded by the Forestry Technology Innovation Program, the Department of Forestry of Guangdong Province (grant number 2018KJCX001, grant number 2017KJCX029).

  1. Author contributions: Conceptualization, Xy.C. and W.Z.; methodology, Xy.C.; validation, Sm.C., Y.R. and Yh.H.; formal analysis, Sm.C.; investigation, Sm.C., Yh.H., Zc.C. and Jt.Z.; resources, W.Z.; data curation, Jj.Z.; writing – original draft preparation, Sm.C.; writing – review and editing, Xy.C.; project administration, W.Z.; funding acquisition, Xy.C.

  2. Conflict of interest: The authors state no conflict of interest.

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Received: 2019-12-25
Revised: 2020-03-22
Accepted: 2020-03-22
Published Online: 2020-06-10

© 2020 Siming Cui et al., published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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  1. Plant Sciences
  2. Dependence of the heterosis effect on genetic distance, determined using various molecular markers
  3. Plant Growth Promoting Rhizobacteria (PGPR) Regulated Phyto and Microbial Beneficial Protein Interactions
  4. Role of strigolactones: Signalling and crosstalk with other phytohormones
  5. An efficient protocol for regenerating shoots from paper mulberry (Broussonetia papyrifera) leaf explants
  6. Functional divergence and adaptive selection of KNOX gene family in plants
  7. In silico identification of Capsicum type III polyketide synthase genes and expression patterns in Capsicum annuum
  8. In vitro induction and characterisation of tetraploid drumstick tree (Moringa oleifera Lam.)
  9. CRISPR/Cas9 or prime editing? – It depends on…
  10. Study on the optimal antagonistic effect of a bacterial complex against Monilinia fructicola in peach
  11. Natural variation in stress response induced by low CO2 in Arabidopsis thaliana
  12. The complete mitogenome sequence of the coral lily (Lilium pumilum) and the Lanzhou lily (Lilium davidii) in China
  13. Ecology and Environmental Sciences
  14. Use of phosphatase and dehydrogenase activities in the assessment of calcium peroxide and citric acid effects in soil contaminated with petrol
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  16. Activity of Vip3Aa1 against Periplaneta americana
  17. Thermostable cellulase biosynthesis from Paenibacillus alvei and its utilization in lactic acid production by simultaneous saccharification and fermentation
  18. Spatiotemporal dynamics of terrestrial invertebrate assemblages in the riparian zone of the Wewe river, Ashanti region, Ghana
  19. Antifungal activity of selected volatile essential oils against Penicillium sp.
  20. Toxic effect of three imidazole ionic liquids on two terrestrial plants
  21. Biosurfactant production by a Bacillus megaterium strain
  22. Distribution and density of Lutraria rhynchaena Jonas, 1844 relate to sediment while reproduction shows multiple peaks per year in Cat Ba-Ha Long Bay, Vietnam
  23. Biomedical Sciences
  24. Treatment of Epilepsy Associated with Common Chromosomal Developmental Diseases
  25. A Mouse Model for Studying Stem Cell Effects on Regeneration of Hair Follicle Outer Root Sheaths
  26. Morphine modulates hippocampal neurogenesis and contextual memory extinction via miR-34c/Notch1 pathway in male ICR mice
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  28. Weight loss may be unrelated to dietary intake in the imiquimod-induced plaque psoriasis mice model
  29. Construction of recombinant lentiviral vector containing human stem cell leukemia gene and its expression in interstitial cells of cajal
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  109. Special Issue On New Approach To Obtain Bioactive Compounds And New Metabolites From Agro-Industrial By-Products
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  114. The impedance analysis of small intestine fusion by pulse source
  115. Errata
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  117. Erratum to “MYL6B drives the capabilities of proliferation, invasion, and migration in rectal adenocarcinoma through the EMT process”
  118. Erratum to “Thermostable cellulase biosynthesis from Paenibacillus alvei and its utilization in lactic acid production by simultaneous saccharification and fermentation”
https://doi.org/10.1515/biol-2020-0034
Keywords for this article
paper mulberry; leaf explant; plant regeneration; explant age; inoculation orientation
Creative Commons
BY 4.0

Articles in the same Issue

  1. Plant Sciences
  2. Dependence of the heterosis effect on genetic distance, determined using various molecular markers
  3. Plant Growth Promoting Rhizobacteria (PGPR) Regulated Phyto and Microbial Beneficial Protein Interactions
  4. Role of strigolactones: Signalling and crosstalk with other phytohormones
  5. An efficient protocol for regenerating shoots from paper mulberry (Broussonetia papyrifera) leaf explants
  6. Functional divergence and adaptive selection of KNOX gene family in plants
  7. In silico identification of Capsicum type III polyketide synthase genes and expression patterns in Capsicum annuum
  8. In vitro induction and characterisation of tetraploid drumstick tree (Moringa oleifera Lam.)
  9. CRISPR/Cas9 or prime editing? – It depends on…
  10. Study on the optimal antagonistic effect of a bacterial complex against Monilinia fructicola in peach
  11. Natural variation in stress response induced by low CO2 in Arabidopsis thaliana
  12. The complete mitogenome sequence of the coral lily (Lilium pumilum) and the Lanzhou lily (Lilium davidii) in China
  13. Ecology and Environmental Sciences
  14. Use of phosphatase and dehydrogenase activities in the assessment of calcium peroxide and citric acid effects in soil contaminated with petrol
  15. Analysis of ethanol dehydration using membrane separation processes
  16. Activity of Vip3Aa1 against Periplaneta americana
  17. Thermostable cellulase biosynthesis from Paenibacillus alvei and its utilization in lactic acid production by simultaneous saccharification and fermentation
  18. Spatiotemporal dynamics of terrestrial invertebrate assemblages in the riparian zone of the Wewe river, Ashanti region, Ghana
  19. Antifungal activity of selected volatile essential oils against Penicillium sp.
  20. Toxic effect of three imidazole ionic liquids on two terrestrial plants
  21. Biosurfactant production by a Bacillus megaterium strain
  22. Distribution and density of Lutraria rhynchaena Jonas, 1844 relate to sediment while reproduction shows multiple peaks per year in Cat Ba-Ha Long Bay, Vietnam
  23. Biomedical Sciences
  24. Treatment of Epilepsy Associated with Common Chromosomal Developmental Diseases
  25. A Mouse Model for Studying Stem Cell Effects on Regeneration of Hair Follicle Outer Root Sheaths
  26. Morphine modulates hippocampal neurogenesis and contextual memory extinction via miR-34c/Notch1 pathway in male ICR mice
  27. Composition, Anticholinesterase and Antipedicular Activities of Satureja capitata L. Volatile Oil
  28. Weight loss may be unrelated to dietary intake in the imiquimod-induced plaque psoriasis mice model
  29. Construction of recombinant lentiviral vector containing human stem cell leukemia gene and its expression in interstitial cells of cajal
  30. Knockdown of lncRNA KCNQ1OT1 inhibits glioma progression by regulating miR-338-3p/RRM2
  31. Protective effect of asiaticoside on radiation-induced proliferation inhibition and DNA damage of fibroblasts and mice death
  32. Prevalence of dyslipidemia in Tibetan monks from Gansu Province, Northwest China
  33. Sevoflurane inhibits proliferation, invasion, but enhances apoptosis of lung cancer cells by Wnt/β-catenin signaling via regulating lncRNA PCAT6/ miR-326 axis
  34. MiR-542-3p suppresses neuroblastoma cell proliferation and invasion by downregulation of KDM1A and ZNF346
  35. Calcium Phosphate Cement Causes Nucleus Pulposus Cell Degeneration Through the ERK Signaling Pathway
  36. Human Dental Pulp Stem Cells Exhibit Osteogenic Differentiation Potential
  37. MiR-489-3p inhibits cell proliferation, migration, and invasion, and induces apoptosis, by targeting the BDNF-mediated PI3K/AKT pathway in glioblastoma
  38. Long non-coding RNA TUG1 knockdown hinders the tumorigenesis of multiple myeloma by regulating the microRNA-34a-5p/NOTCH1 signaling pathway
  39. Large Brunner’s gland adenoma of the duodenum for almost 10 years
  40. Neurotrophin-3 accelerates reendothelialization through inducing EPC mobilization and homing
  41. Hepatoprotective effects of chamazulene against alcohol-induced liver damage by alleviation of oxidative stress in rat models
  42. FXYD6 overexpression in HBV-related hepatocellular carcinoma with cirrhosis
  43. Risk factors for elevated serum colorectal cancer markers in patients with type 2 diabetes mellitus
  44. Effect of hepatic sympathetic nerve removal on energy metabolism in an animal model of cognitive impairment and its relationship to Glut2 expression
  45. Progress in research on the role of fibrinogen in lung cancer
  46. Advanced glycation end product levels were correlated with inflammation and carotid atherosclerosis in type 2 diabetes patients
  47. MiR-223-3p regulates cell viability, migration, invasion, and apoptosis of non-small cell lung cancer cells by targeting RHOB
  48. Knockdown of DDX46 inhibits trophoblast cell proliferation and migration through the PI3K/Akt/mTOR signaling pathway in preeclampsia
  49. Buformin suppresses osteosarcoma via targeting AMPK signaling pathway
  50. Effect of FibroScan test in antiviral therapy for HBV-infected patients with ALT <2 upper limit of normal
  51. LncRNA SNHG15 regulates osteosarcoma progression in vitro and in vivo via sponging miR-346 and regulating TRAF4 expression
  52. LINC00202 promotes retinoblastoma progression by regulating cell proliferation, apoptosis, and aerobic glycolysis through miR-204-5p/HMGCR axis
  53. Coexisting flavonoids and administration route effect on pharmacokinetics of Puerarin in MCAO rats
  54. GeneXpert Technology for the diagnosis of HIV-associated tuberculosis: Is scale-up worth it?
  55. Circ_001569 regulates FLOT2 expression to promote the proliferation, migration, invasion and EMT of osteosarcoma cells through sponging miR-185-5p
  56. Lnc-PICSAR contributes to cisplatin resistance by miR-485-5p/REV3L axis in cutaneous squamous cell carcinoma
  57. BRCA1 subcellular localization regulated by PI3K signaling pathway in triple-negative breast cancer MDA-MB-231 cells and hormone-sensitive T47D cells
  58. MYL6B drives the capabilities of proliferation, invasion, and migration in rectal adenocarcinoma through the EMT process
  59. Inhibition of lncRNA LINC00461/miR-216a/aquaporin 4 pathway suppresses cell proliferation, migration, invasion, and chemoresistance in glioma
  60. Upregulation of miR-150-5p alleviates LPS-induced inflammatory response and apoptosis of RAW264.7 macrophages by targeting Notch1
  61. Long non-coding RNA LINC00704 promotes cell proliferation, migration, and invasion in papillary thyroid carcinoma via miR-204-5p/HMGB1 axis
  62. Neuroanatomy of melanocortin-4 receptor pathway in the mouse brain
  63. Lipopolysaccharides promote pulmonary fibrosis in silicosis through the aggravation of apoptosis and inflammation in alveolar macrophages
  64. Influences of advanced glycosylation end products on the inner blood–retinal barrier in a co-culture cell model in vitro
  65. MiR-4328 inhibits proliferation, metastasis and induces apoptosis in keloid fibroblasts by targeting BCL2 expression
  66. Aberrant expression of microRNA-132-3p and microRNA-146a-5p in Parkinson’s disease patients
  67. Long non-coding RNA SNHG3 accelerates progression in glioma by modulating miR-384/HDGF axis
  68. Long non-coding RNA NEAT1 mediates MPTP/MPP+-induced apoptosis via regulating the miR-124/KLF4 axis in Parkinson’s disease
  69. PCR-detectable Candida DNA exists a short period in the blood of systemic candidiasis murine model
  70. CircHIPK3/miR-381-3p axis modulates proliferation, migration, and glycolysis of lung cancer cells by regulating the AKT/mTOR signaling pathway
  71. Reversine and herbal Xiang–Sha–Liu–Jun–Zi decoction ameliorate thioacetamide-induced hepatic injury by regulating the RelA/NF-κB/caspase signaling pathway
  72. Therapeutic effects of coronary granulocyte colony-stimulating factor on rats with chronic ischemic heart disease
  73. The effects of yam gruel on lowering fasted blood glucose in T2DM rats
  74. Circ_0084043 promotes cell proliferation and glycolysis but blocks cell apoptosis in melanoma via circ_0084043-miR-31-KLF3 axis
  75. CircSAMD4A contributes to cell doxorubicin resistance in osteosarcoma by regulating the miR-218-5p/KLF8 axis
  76. Relationship of FTO gene variations with NAFLD risk in Chinese men
  77. The prognostic and predictive value of platelet parameters in diabetic and nondiabetic patients with sudden sensorineural hearing loss
  78. LncRNA SNHG15 contributes to doxorubicin resistance of osteosarcoma cells through targeting the miR-381-3p/GFRA1 axis
  79. miR-339-3p regulated acute pancreatitis induced by caerulein through targeting TNF receptor-associated factor 3 in AR42J cells
  80. LncRNA RP1-85F18.6 affects osteoblast cells by regulating the cell cycle
  81. MiR-203-3p inhibits the oxidative stress, inflammatory responses and apoptosis of mice podocytes induced by high glucose through regulating Sema3A expression
  82. MiR-30c-5p/ROCK2 axis regulates cell proliferation, apoptosis and EMT via the PI3K/AKT signaling pathway in HG-induced HK-2 cells
  83. CTRP9 protects against MIA-induced inflammation and knee cartilage damage by deactivating the MAPK/NF-κB pathway in rats with osteoarthritis
  84. Relationship between hemodynamic parameters and portal venous pressure in cirrhosis patients with portal hypertension
  85. Long noncoding RNA FTX ameliorates hydrogen peroxide-induced cardiomyocyte injury by regulating the miR-150/KLF13 axis
  86. Ropivacaine inhibits proliferation, migration, and invasion while inducing apoptosis of glioma cells by regulating the SNHG16/miR-424-5p axis
  87. CD11b is involved in coxsackievirus B3-induced viral myocarditis in mice by inducing Th17 cells
  88. Decitabine shows anti-acute myeloid leukemia potential via regulating the miR-212-5p/CCNT2 axis
  89. Testosterone aggravates cerebral vascular injury by reducing plasma HDL levels
  90. Bioengineering and Biotechnology
  91. PL/Vancomycin/Nano-hydroxyapatite Sustained-release Material to Treat Infectious Bone Defect
  92. The thickness of surface grafting layer on bio-materials directly mediates the immuno-reacitivity of macrophages in vitro
  93. Silver nanoparticles: synthesis, characterisation and biomedical applications
  94. Food Science
  95. Bread making potential of Triticum aestivum and Triticum spelta species
  96. Modeling the effect of heat treatment on fatty acid composition in home-made olive oil preparations
  97. Effect of addition of dried potato pulp on selected quality characteristics of shortcrust pastry cookies
  98. Preparation of konjac oligoglucomannans with different molecular weights and their in vitro and in vivo antioxidant activities
  99. Animal Sciences
  100. Changes in the fecal microbiome of the Yangtze finless porpoise during a short-term therapeutic treatment
  101. Agriculture
  102. Influence of inoculation with Lactobacillus on fermentation, production of 1,2-propanediol and 1-propanol as well as Maize silage aerobic stability
  103. Application of extrusion-cooking technology in hatchery waste management
  104. In-field screening for host plant resistance to Delia radicum and Brevicoryne brassicae within selected rapeseed cultivars and new interspecific hybrids
  105. Studying of the promotion mechanism of Bacillus subtilis QM3 on wheat seed germination based on β-amylase
  106. Rapid visual detection of FecB gene expression in sheep
  107. Effects of Bacillus megaterium on growth performance, serum biochemical parameters, antioxidant capacity, and immune function in suckling calves
  108. Effects of center pivot sprinkler fertigation on the yield of continuously cropped soybean
  109. Special Issue On New Approach To Obtain Bioactive Compounds And New Metabolites From Agro-Industrial By-Products
  110. Technological and antioxidant properties of proteins obtained from waste potato juice
  111. The aspects of microbial biomass use in the utilization of selected waste from the agro-food industry
  112. Special Issue on Computing and Artificial Techniques for Life Science Applications - Part I
  113. Automatic detection and segmentation of adenomatous colorectal polyps during colonoscopy using Mask R-CNN
  114. The impedance analysis of small intestine fusion by pulse source
  115. Errata
  116. Erratum to “Diagnostic performance of serum CK-MB, TNF-α and hs-CRP in children with viral myocarditis”
  117. Erratum to “MYL6B drives the capabilities of proliferation, invasion, and migration in rectal adenocarcinoma through the EMT process”
  118. Erratum to “Thermostable cellulase biosynthesis from Paenibacillus alvei and its utilization in lactic acid production by simultaneous saccharification and fermentation”
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