In order to identify the optimum period of spring water-restrictive irrigation for winter wheat (Triticum aestivum L.) in the Haihe Plain, China and elucidate its effects on flag leaf senescence and yield formation, field experiments were conducted at the Xinji Experimental Station of Hebei Agricultural University from 2016 to 2019 by using different irrigation regimes in spring, including the conventional regime involving two irrigation periods (control (CK), the 3-leaf unfolding stage and the anthesis stage) and a series of single, restrictive irrigation regimes (SRI) comprising irrigation at the 3-leaf unfolding stage (3LI), 4LI, 5LI, and 6LI.  There are five major findings: (1) The senescence (determined by the green leaf area, GLA) in the 4LI treatment occurred moderately earlier than that in CK, showed no significant difference with that in 5LI and 6LI, and occurred significantly later than that in 3LI.  (2) Compared with other SRI treatments, the GLA value and photosynthetic rate in 4LI were 14.82 and 20.1% higher, respectively.  Microstructural analysis of flag leaf also revealed that the mesophyll cells and chloroplasts were irregularly arranged under drought stress in 3LI and 6LI; however, drought stress had minimal negative effects on the microstructure in 4LI and 5LI.  (3) Postponed irrigation in spring could significantly increase superoxide dismutase (SOD) and catalase (CAT) activities in the early stage of grain filling; however, these activities would subsequently decrease.  Among the four SRI treatments, the overall enzyme activities were the highest in 4LI, and the combined malondialdehyde (MDA) content in flag leaves in 4LI and 5LI was 14.5% lower on average than that in 3LI and 6LI.  (4) The soluble sugar (SS) and proline (Pro) contents in 4LI were the highest among the four SRI treatments; however, they were lower than those in CK.  The abscisic acid (ABA) hormone content in 4LI and 5LI was lower than that in 3LI and 6LI, respectively, suggesting a smaller drought stress effect in 4LI and 5LI.  (5) In two growing seasons, there was a larger number of spikes per unit area in 4LI (i.e., 13.4% higher than that in 5LI and 6LI) and the 1 000-grain weight in 4LI was the highest among the four SRI treatments (i.e., 6.0% higher than that in the other three SRI treatments).  Therefore, a single restrictive irrigation regime at the 4-leaf unfolding stage is recommended to be effective in slowing down the senescence process of flag leaves and achieving high yield.

 

Somatic embryogenesis (SE) plays a vital role in genetic transformation and massive propagation of important agronomical and economical crops.  Here, we conducted a systematic assessment of the morphological, cytochemical, and cytogenetical characteristics of six culture strains with various embryogenic/regenerative potential during SE process in cotton.  Results indicated that the six cell culture strains had stable ploidy levels, and did not reveal any relationship between the cytogenetic state and their morphogenetic potential.  Moreover, the six culture strains were compared via double staining with Evans blue and Acetocarmine to efficiently distinguish embryogenic and non-embryogenic cells and determine the embryogenic nature of the calli.  In addition, the kind of auxins added in medium affected not only growth property, color, size of cell clumps but also ploidy level and regeneration ability.  By combining analysis of morphological, cytochemical, and cytogenetical characteristics of the cell cultures, we are able to obtain and maintain homogeneous cell population with high morphogenic and regeneration ability and establish efficient somatic embryogenesis and regeneration system from short-term cell cultures in upland cotton, which highlight the application of biotechnological approaches in crop breeding, and above all, to better understand totipotency of cells in higher plants.