RESUMO
Rhizosphere colonization is a pre-requisite for the favorable application of plant growth-promoting rhizobacteria (PGPR). Exchange and mutual recognition of signaling molecules occur frequently between plants and microbes. Here, the luciferase luxAB gene was electrotransformed into the phosphate-solubilizing strain Pseudomonas sp. WS32, a type of plant growth-promoting rhizobacterium with specific affinity for wheat. A labeled WS32 strain (WS32-L) was applied to determine the temporal and spatial traits of colonization within the wheat rhizosphere using rhizoboxes experimentation under natural condition. The effects of colonization on wheat root development and seedling growth were evaluated, and RNA sequencing (RNA-seq) was performed to explore the transcriptional changes that occur in wheat roots under WS32 colonization. The results showed that WS32-L could survive in the wheat rhizosphere for long periods and could expand into new zones following wheat root extension. Significant increases in seedling fresh and dry weight, root fresh and dry weight, root surface area, number of root tips, and phosphorus accumulation in the wheat leaves occurred in response to WS32 rhizosphere colonization. RNA-seq analysis showed that a total of 1485 genes in wheat roots were differentially expressed between the inoculated conditions and the uninoculated conditions. Most of the transcriptional changes occurred for genes annotated to the following functional categories: "phosphorus and other nutrient transport," "hormone metabolism and organic acid secretion," "flavonoid signal recognition," "membrane transport," and "transcription factor regulation." These results are therefore valuable to future studies focused on the molecular mechanisms underlying the growth-promoting activities of PGPR on their host plants.
RESUMO
The basic requirements of plant growth-promoting rhizobacteria (PGPR) for field applications are that they have an affinity for the host plant and that they can colonize the rhizosphere. Here, a new technique was established using soybean agglutin (SBA) as a tool to isolate soybean-specific PGPR. Thirty-three PGPR strains with an affinity for soybean were obtained via the screening method with SBA. All 33 isolates were able to produce indole acetic acid and solubilize inorganic phosphate and potassium. Most isolates (93.94%) were able to solubilize organic phosphate and almost half (45.45%) were able to produce siderophores. More than 40% of the isolates exhibited all five plant growth-promoting traits. The isolate Enterobacter sp. strain DN9 was selected for further analyses of its rhizosphere colonization and soybean growth-promoting effects because of its excellent activity in phosphate and potassium solubilization. The luciferase luxAB gene was electrotransformed into DN9, and the labelled DN9 (DN9-L) was able to survive in the soybean rhizosphere and colonize new spaces as the soybean roots elongated. This strain positively affected root system development and soybean seedling growth. In pot and field experiments, the isolates DN9, DW1, and DW13 significantly increased the nutrient contents in rhizosphere soil and soybean leaves. On average, the seed number per plant and the seed weight per plant were increased by 20% and 24% respectively, in plants inoculated with these PGPR strains in the pot experiment. In a field experiment, compared with uninoculated plants, those inoculated with DW1 showed 46.78% higher pod number per plant and 5.23% higher seed oil content; those inoculated with DW13 showed 79.82% higher seed number per plant and 65.10% higher seed weight per plant; and those inoculated with DN9 showed 9.13% higher 100-seed weight. These results show that SBA can be used as a tool to isolate efficient PGPR to enhance soybean production.
