Type VI protein secretion systems (T6SSs) are specialized transport apparatus which can target both eukaryotic and prokaryotic cells and play key roles in host–pathogen–microbiota interactions. Therefore, T6SSs have attracted much attention as a research topic during the past ten years. In this review, we particularly summarized the T6SS antibacterial function, which involves an interesting offensive and defensive mechanism of the effector–immunity (E–I) pairs. The three main categories of effectors that target the cell wall, membranes, and nucleic acids during bacterial interaction, along with their corresponding immunity proteins are presented. We also discuss structural analyses of several effectors and E–I pairs, which explain the offensive and defensive mechanisms underpinning T6SS function during bacterial competition for niche-space, as well as the bioinformatics, proteomics, and protein–protein interaction (PPI) methods used to identify and characterize T6SS mediated E–I pairs. Additionally, we described PPI methods for verifying E–I pairs.

Proteins are the elementary machinery of life, and their functions are carried out mostly by molecular interactions. Among those interactions, protein–protein interactions (PPIs) are the most important as they participate in or mediate all essential biological processes. However, many common methods for PPI investigations are slightly unreliable and suffer from various limitations, especially in the studies of dynamic PPIs. To solve this problem, a method called Bioluminescence Resonance Energy Transfer (BRET) was developed about seventeen years ago. Since then, BRET has evolved into a whole class of methods that can be used to survey virtually any kinds of PPIs. Compared to many traditional methods, BRET is highly sensitive, reliable, easy to perform, and relatively inexpensive. However, most importantly, it can be done in vivo and allows the real-time monitoring of dynamic PPIs with the easily detectable light signal, which is extremely valuable for the PPI functional research. This review will take a comprehensive look at this powerful technique, including its principles, comparisons with other methods, experimental approaches, classifications, applications, early developments, recent progress, and prospects.

In this study, activated carbon was prepared from Chinese chestnut burs assisted by microwave irradiation with potassium hydroxide (KOH) as activator, and the process conditions were optimized employing Box-Behnken design (BBD) and response surface methodology (RSM). The optimized variables were irradiation time, impregnation time, and mass ratio of alkali-to-carbon, and the iodine adsorption value was used to evaluate the adsorption property of activated carbon. The optimal preparation conditions were determined as follows: irradiation time 17 min, impregnation time 240 min, and mass ratio of alkali-to-char 1.5:1. Meanwhile, the relatively high iodine adsorption value (1141.4 mg/g) was also obtained. Furthermore, the pore structural characterization of activated carbon was analyzed. The analyzed results showed a larger Brunauer-Emmett-Teller (BET) specific surface area (1254.5 m²/g) and a higher microporosity ratio (87.2%), a bigger total pore volume (0.6565 m³/g), but a smaller average pore size (2.093 nm), which demonstrated the obtained activated carbon possessed strong adsorption capacity and well-developed microporous structure. This research could not only establish the foundation of utilizing chestnut burs to prepare activated carbon, but also provide the basis for exploitation of Chinese chestnut by-products. Implications: Because Chinese chestnut burs are the by-products and usually discarded upon harvesting subsequently, the utilization of chestnut burs as a potential source of activated carbon is of great profit to the chestnut processing industries.

Salvia miltiorrhiza Bunge has been widely used in the treatment of cardiovascular and cerebrovascular diseases, due to the pharmacological action of its active components such as the tanshinones. Plasma membrane (PM) H⁺-ATPase plays key roles in numerous physiological processes in plants. However, little is known about the PM H⁺-ATPase gene family in S. miltiorrhiza (Sm). Here, nine PM H⁺-ATPase isoforms were identified and named SmPHA1–SmPHA9. Phylogenetic tree analysis showed that the genetic distance of SmPHAs was relatively far in the S. miltiorrhiza PM H⁺-ATPase family. Moreover, the transmembrane structures were rich in SmPHA protein. In addition, SmPHA4 was found to be highly expressed in roots and flowers. HPLC revealed that accumulation of dihydrotanshinone (DT), cryptotanshinone (CT), and tanshinone I (TI) was significantly reduced in the SmPHA4-OE lines but was increased in the SmPHA4-RNAi lines, ranging from 2.54 to 3.52, 3.77 to 6.33, and 0.35 to 0.74 mg/g, respectively, suggesting that SmPHA4 is a candidate regulator of tanshinone metabolites. Moreover, qRT-PCR confirmed that the expression of tanshinone biosynthetic-related key enzymes was also upregulated in the SmPHA4-RNAi lines. In summary, this study highlighted PM H⁺-ATPase function and provided new insights into regulatory candidate genes for modulating secondary metabolism biosynthesis in S. miltiorrhiza.