Combination of electronically driven micromanipulation with laser desorption ionization mass spectrometry – The unique tool for analysis of seed coat layers and revealing the mystery of seed dormancy

The giant diploid faba genome unlocks variation in a global protein crop

“…Observation and photography were done on an Olympus BX 51 microscope (Olympus) in bright field, and figures were documented with an Apogee U4000 digital camera (Apogee Imaging Systems). For the investigation of metabolite content of surface layers of the hilum by laser desorption–ionization imaging mass spectrometry (LDI-MS), seeds were mechanically cracked and hila with a small part of surrounding tissue were separated from the rest of seed coats using microscissors (MicroSupport), fixed using a double-sided tape on MALDI plates with outer surfaces facing up and analysed as previously described 41 , 112 . LDI-MSI experiments were done using a high-resolution tandem mass spectrometer (HRTMS) Synapt G2-S (Waters).…”

“…Observation and photography were done on an Olympus BX 51 microscope (Olympus) in bright field, and figures were documented with an Apogee U4000 digital camera (Apogee Imaging Systems). For the investigation of metabolite content of surface layers of the hilum by laser desorption-ionization imaging mass spectrometry (LDI-MS), seeds were mechanically cracked and hila with a small part of surrounding tissue were separated from the rest of seed coats using microscissors (MicroSupport), fixed using a double-sided tape on MALDI plates with outer surfaces facing up and analysed as previously described 41,112 16); fresh weights of each tissue were recorded. Because all pods on a given node are not fertilized synchronously and do not necessarily progress through development at the same rate, and on the basis of insights from our previous studies of faba bean seed development 113 , we categorized individual pods into mid and late pod-fill stages in terms of the ratio of cotyledon weight to the total seed weight (Supplementary Fig.…”

Domestication has altered the ABA and gibberellin profiles in developing pea seeds

“…Although wild pea seed dormancy is primarily determined by the water-impermeable seed coat, and its acquisition in legumes is still poorly understood, the elevated ABA level during seed development might be important to prepare the embryo for survival in the dry dormant seed. Similarly to Arabidopsis thaliana seeds, ABA might be involved in the establishment of seed coat impermeability by regulating the synthesis of various compounds such as the hydroxylated fatty acids, phenolics, or pigments (Mendoza et al 2005 ), which were shown to be more abundant in the seed coat of wild peas (Cechová et al 2017 ; Janská et al 2019 ; Krejčí et al 2022 ). Pea seed coat showed abundant accumulation of phenolic compounds that, upon oxidation, may impact seed permeability as well as pigmentation, both typical in the seed coat of dormant peas (Balarynová et al 2022 ; Krejčí et al 2022 ).…”

The giant diploid faba genome unlocks variation in a global protein crop

“…Observation and photography was done on an Olympus BX 51 microscope (Olympus Corp., Tokyo, Japan) in bright field and figures were documented with an Apogee U4000 digital camera (Apogee Imaging Systems, Inc., Roseville, CA, USA). For the investigation of metabolite content of surface layers of the hilum by laser desorption-ionization imaging mass spectrometry (LDI-MS), seeds were mechanically cracked and hila with small part of surrounding tissue were separated from the rest of seed coats using microscissors (MicroSupport, Shizuoka, Japan), fixed using a double-sided tape on MALDI plates with outer surfaces facing up and analysed as previously described 46, 48 . LDI-MSI experiments were done using a high-resolution tandem mass spectrometer (HRTMS) Synapt G2-S (Waters, Milford, USA).…”