The main propagation approach is tissue culture in blueberries, and tissue culture is an effective and low-cost method with higher economic efficiency in blueberries. However, there is a lack of stable and efficient production systems of industrialization of tissue culture in blueberries. In this study, the high-efficiency tissue culture and rapid propagation technology system were established based on blueberry leaves and stems. The optimal medium for callus induction was WPM (woody plant medium) containing 2.0 mg/L Forchlorfenuron (CPPU), 0.2 mg/L 2-isopentenyladenine (2-ip) with a 97% callus induction rate and a callus differentiation rate of 71% by using blueberry leaves as explants. The optimal secondary culture of the leaf callus medium was WPM containing 3.0 mg/L CPPU with an increment coefficient of 24%. The optimal bud growth medium was WPM containing 1.0 mg/L CPPU, 0.4 mg/L 2-ip, with which the growth of the bud was better, stronger and faster. The optimal rooting medium was 1/2 Murashige and Skoog (1/2MS) medium containing 2.0 mg/L naphthylacetic acid (NAA), with which the rooting rate was 90% with shorter rooting time and more adventitious root. In addition, we established a regeneration system based on blueberry stems. The optimal preculture medium in blueberry stem explants was MS medium containing 2-(N-morpholino) ethanesulfonic acid (MES) containing 0.2 mg/L indole-3-acetic acid (IAA), 0.1 mg/L CPPU, 100 mg/L NaCl, with which the germination rate of the bud was 93%. The optimal medium for fast plant growth was MS medium containing MES containing 0.4 mg/L zeatin (ZT), 1 mg/L putrescine, 1 mg/L spermidine, 1 mg/L spermidine, which had a good growth state and growth rate. The optimal cultivation for plantlet growth was MS medium containing MES containing 0.5 mg/L isopentene adenine, with which the plantlet was strong. The optimal rooting medium for the stem was 1/2MS medium containing 2.0 mg/L NAA, with which the rooting rate was 93% with a short time and more adventitious root. In conclusion, we found that stem explants had higher regeneration efficiency for a stable and efficient production system of industrialization of tissue culture. This study provides theoretical guidance and technical support in precision breeding and standardization and industrialization in the blueberry industry.

Blueberry belongs to the Vaccinium genus and is a highly popular fruit crop with significant economic importance. It was not until the early twentieth century that they began to be domesticated through extensive interspecific hybridization. Here, we collected 220 Vaccinium accessions from various geographical locations, including 154 from the United States, 14 from China, eight from Australia, and 29 from Europe and other countries, comprising 164 Vaccinium corymbosum, 15 Vaccinium ashei, 10 lowbush blueberries, seven half-high blueberries, and others. We present the whole-genome variation map of 220 accessions and reconstructed the hundred-year molecular history of interspecific hybridization of blueberry. We focused on the two major blueberry subgroups, the northern highbush blueberry (NHB) and southern highbush blueberry (SHB) and identified candidate genes that contribute to their distinct traits in climate adaptability and fruit quality. Our analysis unveiled the role of gene introgression from Vaccinium darrowii and V. ashei into SHB in driving the differentiation between SHB and NHB, potentially facilitating SHB’s adaptation to subtropical environments. Assisted by genome-wide association studies, our analysis suggested VcTBL44 as a pivotal gene regulator governing fruit firmness in SHB. Additionally, we conducted whole-genome bisulfite sequencing on nine NHB and 12 SHB cultivars, and characterized regions that are differentially methylated between the two subgroups. In particular, we discovered that the β-alanine metabolic pathway genes were enriched for DNA methylation changes. Our study provides high-quality genetic and epigenetic variation maps for blueberry, which offer valuable insights and resources for future blueberry breeding.

Salt is a severe environmental stressor that affects growth and development in plants. It is significant to enhance the salt tolerance in plants. In this study, a salt-responsive WRKY transcription factor PtrWRKY51 was isolated from Populus trichocarpa (clone ‘Nisqually-1′). PtrWRKY51 was highly expressed in mature leaves and root and induced by salt stress. The PtrWRKY51 was overexpressed in Arabidopsis to investigate its biological functions. Compared with Col-0 lines, Overexpressed lines had an increase in germination rate of seed, root length, higher photosynthetic rate, instantaneous leaf WUE, chlorophyll content to improve salt tolerance under salt stress conditions. In contrast, compared to overexpressed and Col-0 lines, the mutant wrky51 was more sensitive to salt stress with lower photosynthetic rate and WUE. Additionally, it was found that the complementary lines (wrky51/ PtrWRKY51) had almost the same salt response as Col-0. In conclusion, PtrWRKY51 is a potential target in the enhancement of poplar tolerance by genetic engineering strategies.

In the present study, the effects of nitrogen (N) supply on water uptake, drought resistance, and hormone regulation were investigated in Populus simonii seedlings grown in hydroponic solution with 5% polyethylene glycol (PEG)-induced drought stress. While acclimating to drought, the P. simonii seedlings exhibited a reduction in growth; differential expression levels of aquaporins (AQPs); activation of auxin (IAA) and abscisic acid (ABA) signaling pathways; a decrease in the net photosynthetic rate and transpiration rate; and an increase in stable nitrogen isotope composition (δ¹⁵N), total soluble substances, and intrinsic water use efficiency (WUEi), with a shift in the homeostasis of reactive oxygen species (ROS) production and scavenging. A low N supply (0.01 mM NH₄NO₃) or sufficient N supply (1 mM NH₄NO₃) exhibited distinct morphological, physiological, and transcriptional responses during acclimation to drought, primarily due to strong responses in the transcriptional regulation of genes encoding AQPs; higher soluble phenolics, total N concentrations, and ROS scavenging; and lower transpiration rates, IAA content, ABA content, and ROS accumulation with a sufficient N supply. P. simonii can differentially manage water uptake and hormone modulation in response to drought stress under deficient and sufficient N conditions. These results suggested that increased N may contribute to drought tolerance by decreasing the transpiration rate and O₂⁻ production while increasing water uptake and antioxidant enzyme activity.